Accurate analyte measurements for electrochemical test strip based on sensed physical characteristic(s) of the sample containing the analyte

ABSTRACT

Various embodiments for methods and systems that allow for a more accurate analyte concentration with a biosensor by determining at least one physical characteristic of the sample containing the analyte and deriving one of a batch slope, sampling time, or combinations thereof to attain accurate glucose concentration.

PRIORITY

This DIVISIONAL application claims the benefits of priority under 35 USC§§ 120 and 121 from prior filed U.S. application Ser. No. 14/353,870filed on Apr. 24, 2014, allowed, which prior filed application (Ser. No.14/353,870) is a National Stage application of International ApplicationPCT/GB2012/053276 filed on Dec. 28, 2012 claims the benefits of priorityof prior International Patent Application PCT/GB2012/053277 andPCT/GB2012/053279 both filed on Dec. 28, 2012, in which each of thereferenced herein International Patent Applications claims benefits ofpriority to U.S. Provisional Patent Application Ser. Nos. 61/581,087filed Dec. 29, 2011; 61/581,089 filed Dec. 29, 2011; 61/581,099 filedDec. 29, 2011; and 61/581,100 filed Dec. 29, 2011; and U.S. ProvisionalPatent Application Ser. No. 61/654,013 filed May 31, 2012, in which allthe prior patent applications referenced herein are hereby incorporatedby reference as if fully set forth herein for this application.

BACKGROUND

Electrochemical glucose test strips, such as those used in the OneTouch®Ultra® whole blood testing kit, which is available from LifeScan, Inc.,are designed to measure the concentration of glucose in a physiologicalfluid sample from patients with diabetes. The measurement of glucose canbe based on the selective oxidation of glucose by the enzyme glucoseoxidase (GO). The reactions that can occur in a glucose test strip aresummarized below in Equations 1 and 2.Glucose+GO_((ox))→Gluconic Acid+GO_((red))  Eq. 1GO_((red))+2Fe(CN)₆ ³⁻→GO_((ox))+2Fe(CN)₆ ⁴⁻  Eq. 2

As illustrated in Equation 1, glucose is oxidized to gluconic acid bythe oxidized form of glucose oxidase (GO_((ox))). It should be notedthat GO_((ox)) may also be referred to as an “oxidized enzyme.” Duringthe reaction in Equation 1, the oxidized enzyme GO_((ox)) is convertedto its reduced state, which is denoted as GO_((red)) (i.e., “reducedenzyme”). Next, the reduced enzyme GO_((red)) is re-oxidized back toGO_((ox)) by reaction with Fe(CN)₆ ³⁻ (referred to as either theoxidized mediator or ferricyanide) as illustrated in Equation 2. Duringthe re-generation of GO_((red)) back to its oxidized state GO_((ox)),Fe(CN)₆ ³⁻ is reduced to Fe(CN)₆ ⁴⁻ (referred to as either reducedmediator or ferrocyanide).

When the reactions set forth above are conducted with a test signalapplied between two electrodes, a test current can be created by theelectrochemical re-oxidation of the reduced mediator at the electrodesurface. Thus, since, in an ideal environment, the amount offerrocyanide created during the chemical reaction described above isdirectly proportional to the amount of glucose in the sample positionedbetween the electrodes, the test current generated would be proportionalto the glucose content of the sample. A mediator, such as ferricyanide,is a compound that accepts electrons from an enzyme such as glucoseoxidase and then donates the electrons to an electrode. As theconcentration of glucose in the sample increases, the amount of reducedmediator formed also increases; hence, there is a direct relationshipbetween the test current, resulting from the re-oxidation of reducedmediator, and glucose concentration. In particular, the transfer ofelectrons across the electrical interface results in the flow of a testcurrent (2 moles of electrons for every mole of glucose that isoxidized). The test current resulting from the introduction of glucosecan, therefore, be referred to as a glucose signal.

Electrochemical biosensors may be adversely affected by the presence ofcertain blood components that may undesirably affect the measurement andlead to inaccuracies in the detected signal. This inaccuracy may resultin an inaccurate glucose reading, leaving the patient unaware of apotentially dangerous blood sugar level, for example. As one example,the blood hematocrit level (i.e. the percentage of the amount of bloodthat is occupied by red blood cells) can erroneously affect a resultinganalyte concentration measurement.

Variations in a volume of red blood cells within blood can causevariations in glucose readings measured with disposable electrochemicaltest strips. Typically, a negative bias (i.e., lower calculated analyteconcentration) is observed at high hematocrit, while a positive bias(i.e., higher calculated analyte concentration) is observed at lowhematocrit. At high hematocrit, for example, the red blood cells mayimpede the reaction of enzymes and electrochemical mediators, reduce therate of chemistry dissolution since there is less plasma volume tosolvate the chemical reactants, and slow diffusion of the mediator.These factors can result in a lower than expected glucose reading asless signal is produced during the electrochemical process. Conversely,at low hematocrit, fewer red blood cells may affect the electrochemicalreaction than expected, and a higher measured signal can result. Inaddition, the physiological fluid sample resistance is also hematocritdependent, which can affect voltage and/or current measurements.

Several strategies have been used to reduce or avoid hematocrit basedvariations on blood glucose. For example, test strips have been designedto incorporate meshes to remove red blood cells from the samples, orhave included various compounds or formulations designed to increase theviscosity of red blood cells and attenuate the effect of low hematocriton concentration determinations. Other test strips have included lysisagents and systems configured to determine hemoglobin concentration inan attempt to correct hematocrit. Further, biosensors have beenconfigured to measure hematocrit by measuring an electrical response ofthe fluid sample via alternating current signals or change in opticalvariations after irradiating the physiological fluid sample with light,or measuring hematocrit based on a function of sample chamber fill time.These sensors have certain disadvantages. A common technique of thestrategies involving detection of hematocrit is to use the measuredhematocrit value to correct or change the measured analyteconcentration, which technique is generally shown and described in thefollowing respective US Patent Application Publication Nos.2010/0283488; 2010/0206749; 2009/0236237; 2010/0276303; 2010/0206749;2009/0223834; 2008/0083618; 2004/0079652; 2010/0283488; 2010/0206749;2009/0194432; or U.S. Pat. Nos. 7,972,861 and 7,258,769, all of whichare incorporated by reference herein to this application.

SUMMARY OF THE DISCLOSURE

Applicants have provided various embodiments of a technique to allow forimproved glucose measurement using a relationship between batch slopeand physical characteristic (e.g., hematocrit) to derive a new batchslope that can be used to determine the analyte concentration based onthis derived batch slope of an electrochemical biosensor.Advantageously, this new technique does not rely on correction(s) ormodification(s) to be made to an analyte measurement, thereby reducingtest time while at the same time improving accuracy.

In a first aspect of applicants' disclosure, a method of determining ananalyte concentration from a fluid sample (which may be a physiologicalsample) with a biosensor (which may be in the form of a test strip butis not limited to a test strip) is provided. The biosensor has at leasttwo electrodes and a reagent disposed on at least one of the electrodes.The method can be achieved by: depositing a fluid sample (which may be aphysiological sample) on the at least two electrodes to start an analytetest sequence; applying a first signal to the sample to measure orestimate a physical characteristic of the sample; deriving a batch slopefor the biosensor based on the measured or estimated physicalcharacteristic from an equation of the form:x=aH ² +bH+cwhere

x represents a derived batch slope;

H is measured or estimated physical characteristic;

a represents about 1.4e-6, or is equal to 1.4e-6, or is equal to1.4e-6+/−10%, 5% or 1% of 1.4e-6;

b represents about −3.8e-4, or is equal to −3.8e-4, or is equal to−3.8e-4+/−10%, 5% or 1% of −3.8e-4;

c represents about 3.6e-2, or is equal to 3.6e-2, or is equal to−3.6e-2+/−10%, 5% or 1% of 3.6e-2;

driving a second signal to the sample; and measuring an output signalfrom at least one of the at least two electrodes;

calculating an analyte concentration based on the measured output signaland derived batch slope with an equation of the form:

$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{x} \right\rbrack$where

G₀ represents an analyte concentration

I_(E) represents, or is, a signal (value or measurement; proportional toanalyte concentration) measured at a predetermined or specified samplingtime, which may be the total signal measured at the predetermined orspecified sampling time;

Intercept represents a calibration parameter for a batch of biosensors;

x represents, or is, the derived batch slope from the deriving step. Theterm “e”, as used throughout this disclosure, refers to scientificnotation. For example, 1.4 e-6 is 1.4×10⁻⁶ or 0.0000014.

In a second aspect of applicants' disclosure, a method of determining ananalyte concentration from a fluid sample (which may be a physiologicalsample) with a biosensor (which may be in the form of a test strip butis not limited to a test strip) is provided. The biosensor has at leasttwo electrodes and a reagent disposed on at least one of the electrodes.The method can be achieved by: depositing a fluid sample (which may be aphysiological sample) on the at least two electrodes to start an analytetest sequence; applying a first signal to the sample to measure aphysical characteristic of the sample; deriving a batch slope for thebiosensor based on the measured or estimated physical characteristic;driving a second signal to the sample; and measuring an output signalfrom at least one of the at least two electrodes; calculating an analyteconcentration based on the measured output signal and derived batchslope from the measured or estimated physical characteristic of thesample.

In any of the aspects described herein the following features may alsobe utilized in various combinations with the previously disclosedaspects: the applying of the first signal and the driving of the secondsignal may be in sequential order; the applying of the first signal mayoverlap with the driving of the second signal; the applying of the firstsignal may include directing an alternating signal to the sample so thata physical characteristic of the sample is determined from an output ofthe alternating signal; the applying of the first signal may includedirecting an optical signal to the sample so that a physicalcharacteristic of the sample is determined from an output of the opticalsignal; the physical characteristic may include hematocrit and theanalyte may include glucose; the physical characteristic may include atleast one of viscosity, hematocrit, temperature and density of thesample; the directing may include driving first and second alternatingsignals at different respective frequencies in which a first frequencyis lower than the second frequency; the first frequency may be at leastone order of magnitude lower than the second frequency; and/or the firstfrequency may include any frequency in the range of about 10 kHz toabout 250 kHz, or about 10 kHz to about 90 kHz.

In these aspects, the deriving may include calculating a batch slopefrom an equation of the form:x=aH ² +bH+cwhere

x represents, or is, a derived batch slope from the deriving step;

H represents, or is, measured or estimated physical characteristic (e.g.hematocrit);

a represents about 1.4e-6, or is equal to 1.4e-6, or is equal to1.4e-6+/−10%, 5% or 1% of the numerical value provided hereof;

b represents about −3.8e-4, or is equal to −3.8e-4, or is equal to−3.8e-4+/−10%, 5% or 1% of the numerical value provided hereof;

c represents about 3.6e-2, or is equal to 3.6e-2, or is equal to−3.6e-2+/−10%, 5% or 1% of the numerical value provided hereof. The term“e”, as used throughout this disclosure, refers to scientific notation.For example, 1.4 e-6 is 1.4×10⁻⁶ or 0.0000014.

Furthermore, the calculating of the analyte concentration may includeutilizing an equation of the form:

$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{x} \right\rbrack$where

G₀ represents, or is, an analyte concentration

I_(E) represents, or is, a signal (proportional to analyteconcentration) which may be the total signal measured at a predeterminedtime, for example at least one of 2.5 seconds or 5 seconds, or at one of2.5 seconds or 5 seconds (about or exactly) after a start of the testsequence; and wherein the total signal means one signal from eachelectrode summed or the signal from one electrode being doubled;

Intercept represents, or is, a calibration parameter for a batch ofbiosensors;

x represents, or is, a derived batch slope from the deriving step.

In a third aspect of applicants' disclosure, an analyte measurementsystem is provided that includes a test strip and an analyte meter. Thetest strip includes a substrate with a plurality of electrodes connectedto respective electrode connectors. The analyte meter includes ahousing, a test strip port, and a processor. The strip port connector isconfigured to connect to the respective electrode connectors of the teststrip. The microprocessor is in electrical communication with the teststrip port connector to apply electrical signals or sense electricalsignals from the plurality of electrodes during a test sequence. Themicroprocessor is configured to, during the test sequence: (a) apply afirst signal to the plurality of electrodes so that a batch slopedefined by a physical characteristic of a physiological fluid sample isderived and (b) apply a second signal to the plurality of electrodes sothat an analyte concentration is determined based on the derived batchslope.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: theplurality of electrodes may include at least two electrodes to measurethe physical characteristic and at least two other electrodes to measurethe analyte concentration; the at least two electrodes and the at leasttwo other electrodes may be disposed in the same chamber provided on thesubstrate; the at least two electrodes and the at least two otherelectrodes may be disposed in different chambers provided on thesubstrate; the at least two electrodes may include two electrodes tomeasure the physical characteristic and the analyte concentration; theplurality of electrodes may include two electrodes to measure thephysical characteristic and the analyte concentration; alternatively,all of the electrodes are disposed on the same plane defined by thesubstrate; a reagent is disposed proximate the at least two otherelectrodes and no reagent is disposed on the at least two electrodes;and/or the batch slope may be calculated from an equation of the form:x=aH ² +bH+cwhere

x represents, or is, a derived batch slope from the deriving step;

H represents, or is, measured or estimated physical characteristic (e.g.hematocrit);

a represents about 1.4e-6, or is equal to 1.4e-6, or is equal to1.4e-6+/−10%, 5% or 1% of the numerical value provided hereof;

b represents about −3.8e-4, or is equal to −3.8e-4, or is equal to−3.8e-4+/−10%, 5% or 1% of the numerical value provided hereof;

c represents about 3.6e-2, or is equal to 3.6e-2, or is equal to−3.6e-2+/−10%, 5% or 1% of the numerical value provided herein. The term“e”, as used throughout this disclosure, refers to scientific notation.Accordingly, 1.4 e-6 is 1.4×10⁻⁶ or 0.0000014.

Furthermore, in these aspects, the analyte concentration may bedetermined from an equation of the form:

$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{x} \right\rbrack$where

G₀ represents, or is, an analyte concentration

I_(E) represents, or is, a signal (proportional to analyteconcentration) measured at a predetermined or specified sampling time,which may be the total signal measured at the predetermined or specifiedsampling time;

Intercept represents, or is, the calibration parameter for a batch oftest strips;

x represents, or is, the derived batch slope from the deriving step.

In a fourth aspect of applicants' disclosure, an analyte measurementsystem is provided that includes a test strip and an analyte meter. Thetest strip includes a substrate with a plurality of electrodes connectedto respective electrode connectors. The analyte meter includes ahousing, a test strip port, and a processor. The strip port connector isconfigured to connect to the respective electrode connectors of the teststrip. The microprocessor is in electrical communication with the teststrip port connector to apply electrical signals or sense electricalsignals from the plurality of electrodes during a test sequence. Themicroprocessor is configured to, during a test sequence: (a) apply afirst signal to the plurality of electrodes so that a batch slopedefined by a physical characteristic of a physiological fluid sample isderived and (b) apply a second signal to the plurality of electrodes sothat an analyte concentration is determined based on the derived batchslope obtained from the physical characteristic of the sample withinabout 10 seconds of a start of the test sequence.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: theplurality of electrodes may include at least two electrodes to measurethe physical characteristic and at least two other electrodes to measurethe analyte concentration; the at least two electrodes and the at leasttwo other electrodes may be disposed in the same chamber provided on thesubstrate; the at least two electrodes and the at least two otherelectrodes may be disposed in different chambers provided on thesubstrate; the at least two electrodes may include two electrodes tomeasure the physical characteristic and the analyte concentration; theplurality of electrodes may include two electrodes to measure thephysical characteristic and the analyte concentration; all of theelectrodes may be disposed on the same plane defined by the substrate; areagent is disposed proximate the at least two other electrodes and noreagent disposed on the at least two electrodes; and/or the batch slopemay be calculated from an equation of the form:x=aH ² +bH+cwhere

x represents, or is, a derived batch slope from the deriving step;

H represents, or is, measured or estimated physical characteristic (e.g.hematocrit);

a represents about 1.4e-6, or is equal to 1.4e-6, or is equal to1.4e-6+/−10%, 5% or 1% of the numerical value provided hereof;

b represents about −3.8e-4, or is equal to −3.8e-4, or is equal to−3.8e-4+/−10%, 5% or 1% of the numerical value provided hereof;

c represents about 3.6e-2, or is equal to 3.6e-2, or is equal to−3.6e-2+/−10%, 5% or 1% of the numerical value provided hereof. The term“e”, as used throughout this disclosure, refers to scientific notation.Accordingly, 1.4 e-6 is 1.4×10⁻⁶ or 0.0000014.

In these aspects, the analyte concentration may be calculated from anequation of the form:

$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{x} \right\rbrack$where

G₀ represents, or is, an analyte concentration

I_(E) represents, or is, a signal (value or measurement; proportional toanalyte concentration) measured at a predetermined or specified samplingtime, which may be the total signal measured at the predetermined orspecified sampling time;

Intercept represents, or is, a calibration parameter for a batch of teststrips;

x represents, or is, a derived batch slope from the deriving step.

In a fifth aspect of applicants' disclosure, a method of demonstratingincreased accuracy of a test strip is provided. The method can beachieved by: providing a batch of test strips; introducing a referentialsample containing a referential concentration of an analyte to each teststrip of the batch of test strips to initiate a test sequence; reactingthe analyte with a reagent on each test strip to cause a physicaltransformation of the analyte proximate the two electrodes (which may bebetween the two electrodes); determining a physical characteristic ofthe referential sample; deriving a batch slope for the batch of teststrips based on the determined physical characteristic of thereferential sample; sampling an electrical output of the referentialsample at a predetermined time point during the test sequence;calculating an analyte concentration based on the defined batch slopeand sampled electrical output to provide for a final analyteconcentration value for each test strip of the batch of test strips suchthat at least 95% of the final analyte concentration values of the batchof test strips are within ±15% of the referential analyte concentration.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: thedetermining may include applying a first signal to the sample to measurea physical characteristic of the sample; the sampling may includedriving a second signal to the sample; the applying of the first signaland the driving of the second signal may be in sequential order; theapplying of the first signal may overlap with the driving of the secondsignal; the applying of the first signal may include directing analternating signal to the sample so that a physical characteristic ofthe sample is determined from an output of the alternating signal; theapplying of the first signal may include directing an optical signal tothe sample so that a physical characteristic of the sample is determinedfrom an output of the optical signal; the physical characteristic mayinclude hematocrit and the analyte may include glucose; the physicalcharacteristic may include at least one of viscosity, hematocrit,temperature and density; the directing may include driving first andsecond alternating signals at different respective frequencies in whicha first frequency is lower than the second frequency; the firstfrequency may be at least one order of magnitude lower than the secondfrequency; the first frequency may include any frequency in the range ofabout 10 kHz to about 250 kHz, or about 10 kHz to about 90 kHz; and/orthe deriving may include calculating a batch slope from an equation ofthe form:x=aH ² +bH+cwhere

x represents, or is, a derived batch slope from the deriving step;

H represents, or is, measured, determined or estimated physicalcharacteristic (e.g. hematocrit);

a represents about 1.4e-6, or is equal to 1.4e-6, or is equal to1.4e-6+/−10%, 5% or 1% of the numerical value provided hereof;

b represents about −3.8e-4, or is equal to −3.8e-4, or is equal to−3.8e-4+/−10%, 5% or 1% of the numerical value provided hereof;

c represents about 3.6e-2, or is equal to 3.6e-2, or is equal to−3.6e-2+/−10%, 5% or 1% of the numerical value provided hereof. The term“e”, as used throughout this disclosure, refers to scientific notation.Accordingly, 1.4 e-6 is 1.4×10⁻⁶ or 0.0000014.

In these aspects, the calculating of the analyte concentration mayinclude utilizing an equation of the form:

$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{x} \right\rbrack$where

G₀ represents, or is, an analyte concentration

I_(E) represents, or is, a signal (value or measurement; proportional toanalyte concentration) measured at a predetermined or specified samplingtime, which may be the total signal measured at the predetermined orspecified sampling time;

Intercept represents, or is, a calibration parameter for a batch of teststrips;

x represents, or is, a derived batch slope from the deriving step.

In a sixth aspect of applicants' disclosure, a method of determining ananalyte concentration from a fluid sample (which may be a physiologicalsample) is provided. The method can be achieved by: depositing a fluidsample (which may be a physiological sample) on a biosensor; applyingsignals to the sample to transform the analyte into a differentmaterial; measuring or estimating a physical characteristic of thesample; evaluating signal output from the sample; deriving a parameterof the biosensor from the measured or estimated physical characteristic;and determining an analyte concentration based on the derived parameterof the biosensor and the signal output of the sample.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: themeasuring may include applying a first signal to the sample to measure aphysical characteristic of the sample; the evaluating may includedriving a second signal to the sample; the applying of the first signaland the driving of the second signal may be in sequential order; theapplying of the first signal may overlap with the driving of the secondsignal; the applying of the first signal may include directing analternating signal to the sample so that a physical characteristic ofthe sample is determined from an output of the alternating signal; theapplying of the first signal may include directing an optical signal tothe sample so that a physical characteristic of the sample is determinedfrom an output of the optical signal; the physical characteristic mayinclude hematocrit and the analyte may include glucose; the physicalcharacteristic may include at least one of viscosity, hematocrit,temperature and density; the directing may include driving first andsecond alternating signals at different respective frequencies in whicha first frequency is lower than the second frequency; the firstfrequency may be at least one order of magnitude lower than the secondfrequency; the first frequency may include any frequency in the range ofabout 10 kHz to about 250 kHz, or about 10 kHz to about 90 kHz; thederived parameter may be a batch slope; and/or the deriving may includecalculating a batch slope from an equation of the form:x=aH ² +bH+cwhere

x represents, or is, a derived batch slope from the deriving step;

H represents, or is, measured or estimated physical characteristic (e.g.hematocrit);

a represents about 1.4e-6, or is equal to 1.4e-6, or is equal to1.4e-6+/−10%, 5% or 1% of the numerical value provided hereof;

b represents about −3.8e-4, or is equal to −3.8e-4, or is equal to−3.8e-4+/−10%, 5% or 1% of the numerical value provided hereof;

c represents about 3.6e-2, or is equal to 3.6e-2, or is equal to−3.6e-2+/−10%, 5% or 1% of the numerical value provided herein. The term“e”, as used throughout this disclosure, refers to scientific notation.Accordingly, 1.4 e-6 is 1.4×10⁻⁶ or 0.0000014.

In these aspects, the calculating of the analyte concentration mayinclude utilizing an equation of the form:

$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{x} \right\rbrack$where

G₀ represents, or is, an analyte concentration

I_(E) represents, or is, a signal (value or measurement; proportional toanalyte concentration) measured at a predetermined or specified samplingtime, which may be the total signal measured at the predetermined orspecified sampling time;

Intercept represents, or is, a calibration parameter for a batch of teststrips;

x represents a derived batch slope from the deriving step.

In a seventh aspect of applicants' disclosure, a method of determiningan analyte concentration from a fluid sample (which may be aphysiological sample) with a biosensor (e.g. a test strip) is provided.The biosensor has at least two electrodes and a reagent disposed on atleast one electrode of the electrodes. The method can be achieved by:depositing a fluid sample (which may be a physiological sample) on theat least two electrodes to start an analyte test sequence; applying afirst signal to the sample to derive a physical characteristic of thesample; obtaining a physical characteristic of the sample; specifying asampling time based on the obtained physical characteristic; driving asecond signal to the sample; and measuring an output signal at thespecified sampling time from at least one electrode of the at least twoelectrodes; and calculating an analyte concentration based on themeasured output signal.

For the seventh aspect of applicants' disclosure, the following featuresmay also be utilized in various combinations: the applying of the firstsignal and the driving of the second signal may be in sequential order;the applying of the first signal may overlap with the driving of thesecond signal; the applying of the first signal may include directing analternating signal to the sample so that a physical characteristic ofthe sample is determined from an output of the alternating signal; theapplying of the first signal may include directing an optical signal tothe sample so that a physical characteristic of the sample is determinedfrom an output of the optical signal; the physical characteristic mayinclude hematocrit and the analyte may include glucose; the physicalcharacteristic may include at least one of viscosity, hematocrit,temperature and density of the sample; the directing may include drivingfirst and second alternating signals at different respective frequenciesin which a first frequency is lower than the second frequency; the firstfrequency may be at least one order of magnitude lower than the secondfrequency; the first frequency may include any frequency in the range ofabout 10 kHz to about 250 kHz, or about 10 kHz to about 90 kHz; and/orthe specified sampling time may be calculated using an equation of theform:SpecifiedSamplingTime=x ₁ H ^(x2) +x ₃where

“SpecifiedSamplingTime” is designated as a time point from the start ofthe test sequence at which to sample or measure the output signal (e.g.output signal) of the test strip,

H represents the physical characteristic of the sample;

x₁ is about 4.3e5, or is equal to 4.3e5, or is equal to 4.3e5+/−10%, 5%or 1% of the numerical value provided hereof;

x₂ is about −3.9, or is equal to −3.9, or is equal to −3.9+/−10%, 5% or1% of the numerical value provided hereof; and

x₃ is about 4.8, or is equal to 4.8, or is equal to 4.8+/−10%, 5% or 1%of the numerical value provided herein. The term “e”, as used throughoutthis disclosure, refers to scientific notation. Accordingly, 4.3e5 is4.3×10⁵ or 430,000.

Additionally, in the seventh aspect of applicants' disclosure, thecalculating of the analyte concentration may be computed with anequation of the form:

$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{Slope} \right\rbrack$where

G₀ represents, or is, an analyte concentration

I_(E) represents, or is, a signal (proportional to analyteconcentration) measured at the SpecifiedSamplingTime, which may be thetotal signal measured at the SpecifiedSamplingTime;

Slope represents, or is, the value obtained from calibration testing ofa batch of test strips of which this particular strip comes from; and

Intercept represents, or is, the value obtained from calibration testingof a batch of test strips from which this particular strip comes.

In an eighth aspect of applicants' disclosure, an analyte measurementsystem is provided that includes a test strip and a test meter. The teststrip includes a substrate and a plurality of electrodes connected torespective electrode connectors. The analyte meter includes a housing, atest strip port connector configured to connect to the respectiveelectrode connectors of the test strip and a microprocessor. Themicroprocessor is in electrical communication with the test strip portconnector to apply electrical signals or sense electrical signals fromthe plurality of electrodes during a test sequence. The microprocessoris configured to, during the test sequence: (a) apply a first signal tothe plurality of electrodes so that a specific sampling time pointdetermined from a physical characteristic of a physiological fluidsample is derived, (b) apply a second signal to the plurality ofelectrodes, and (c) measure a signal output from one of the plurality ofelectrodes at the specified sampling time so that an analyteconcentration is determined.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: theplurality of electrodes may include at least two electrodes to measurethe physical characteristic and at least two other electrodes to measurethe analyte concentration; the at least two electrodes and the at leasttwo other electrodes may be disposed in the same chamber provided on thesubstrate; the at least two electrodes and the at least two otherelectrodes may be disposed in different chambers provided on thesubstrate; the at least two electrodes may comprise two electrodes tomeasure the physical characteristic and the analyte concentration; theplurality of electrodes may comprise two electrodes to measure thephysical characteristic and the analyte concentration; all of theelectrodes may be disposed on the same plane defined by the substrate; areagent may be disposed proximate the at least two other electrodes andno reagent disposed on the at least two electrodes; and/or the specifiedsampling time may be calculated using an equation of the form:SpecifiedSamplingTime=x ₁ H ^(x) ² +x ₃where

“SpecifiedSamplingTime” is designated as a time point from the start ofthe test sequence at which to sample the output signal (e.g. outputsignal) of the test strip,

H represents the physical characteristic of the sample;

x₁ is about 4.3e5, or is equal to 4.3e5, or is equal to 4.3e5+/−10%, 5%or 1% of the numerical value provided hereof;

x₂ is about −3.9, or is equal to −3.9, or is equal to −3.9+/−10%, 5% or1% of the numerical value provided hereof; and

x₃ is about 4.8, or is equal to 4.8, or is equal to 4.8+/−10%, 5% or 1%of the numerical value provided herein. The term “e”, as used throughoutthis disclosure, refers to scientific notation. Accordingly, 4.3e5 is4.3×10⁵ or 430,000.

Additionally, in these aspects, the analyte concentration is determinedfrom an equation of the form:

$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{Slope} \right\rbrack$where

G₀ represents, or is, an analyte concentration

I_(E) represents, or is, a signal (value or measurement; proportional toanalyte concentration) measured at the SpecifiedSamplingTime, which maybe the total signal measured at the SpecifiedSamplingTime;

Slope represents, or is, the value obtained from calibration testing ofa batch of test strips of which this particular strip comes from; and

Intercept represents, or is, the value obtained from calibration testingof a batch of test strips from which this particular strip comes from.

In a ninth aspect of applicants' disclosure, an analyte measurementsystem is provided that includes a test strip and a test meter. The teststrip includes a substrate and a plurality of electrodes connected torespective electrode connectors. The analyte meter includes a housing, atest strip port connector configured to connect to the respectiveelectrode connectors of the test strip and a microprocessor. Themicroprocessor is in electrical communication with the test strip portconnector to apply electrical signals or sense electrical signals fromthe plurality of electrodes. The microprocessor is configured to, duringa test sequence: (a) apply a first signal to the plurality of electrodesso that a specified sampling time determined from a physicalcharacteristic of a physiological fluid sample is derived, (b) apply asecond signal to the plurality of electrodes, and (c) measure a signaloutput from one of the plurality of electrodes at the specified samplingtime so that an analyte concentration of the sample is determined basedon the specified sampling time within about 10 seconds of a start of thetest sequence.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: theplurality of electrodes may include at least two electrodes to measurethe physical characteristic and at least two other electrodes to measurethe analyte concentration; the at least two electrodes and the at leasttwo other electrodes may be disposed in the same chamber provided on thesubstrate; the at least two electrodes and the at least two otherelectrodes may be disposed in different chambers provided on thesubstrate; the at least two electrodes may comprise two electrodes tomeasure the physical characteristic and the analyte concentration; allof the electrodes may be disposed on the same plane defined by thesubstrate; a reagent may be disposed proximate the at least two otherelectrodes and no reagent disposed on the at least two electrodes;and/or the specified sampling time may be calculated using an equationof the form:SpecifiedSamplingTime=x ₁ H ^(x) ² +x ₃where

“SpecifiedSamplingTime” is designated as a time point from the start ofthe test sequence at which to sample the output signal (e.g. outputsignal) of the test strip,

H represents, or is, the physical characteristic of the sample;

x₁ is about 4.3e5, or is equal to 4.3e5, or is equal to 4.3e5+/−10%, 5%or 1% of the numerical value provided hereof;

x₂ is about −3.9, or is equal to −3.9, or is equal to −3.9+/−10%, 5% or1% of the numerical value provided hereof; and

x₃ is about 4.8, or is equal to 4.8, or is equal to 4.8+/−10%, 5% or 1%of the numerical value provided herein. The term “e”, as used throughoutthis disclosure, refers to scientific notation. Accordingly, 4.3e5 is4.3×10⁵ or 430,000.

Additionally, for these aspects, an analyte concentration may becalculated from an equation of the form:

$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{Slope} \right\rbrack$where

G₀ represents, or is, an analyte concentration

I_(E) represents, or is, a signal (proportional to analyteconcentration) measured at the SpecifiedSamplingTime, which may be thetotal signal measured at the SpecifiedSamplingTime;

Slope represents, or is, the value obtained from calibration testing ofa batch of test strips of which this particular strip comes from; and

Intercept represents, or is, the value obtained from calibration testingof a batch of test strips of which this particular strip comes from.

In a tenth aspect of applicants' disclosure, a method of determining ananalyte concentration from a fluid sample (which may be a physiologicalsample) is provided. The method can be achieved by: depositing a fluidsample (which may be a physiological sample) on a biosensor (e.g. a teststrip) having a reagent deposited thereon; applying signals to thesample and the reagent to transform the analyte into a differentmaterial; obtaining a physical characteristic of the sample; specifyinga time point for sampling of signal output based on the obtainedphysical characteristic; measuring signal output at the specifiedsampling time; and determining an analyte concentration based on themeasured signal output of the sample.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: theobtaining may include driving a second signal to the sample to derive aphysical characteristic of the sample; the applying may include applyinga first signal to the sample to derive a physical characteristic of thesample, and the applying of the first signal and the driving of thesecond signal may be in sequential order; the applying of the firstsignal may overlap with the driving of the second signal; the applyingmay comprise applying a first signal to the sample to derive a physicalcharacteristic of the sample, and the applying of the first signal mayoverlap with the driving of the second signal; the applying of the firstsignal may include directing an alternating signal to the sample so thata physical characteristic of the sample is determined from an output ofthe alternating signal; the applying of the first signal may includedirecting an optical signal to the sample so that a physicalcharacteristic of the sample is determined from an output of the opticalsignal; the physical characteristic may include hematocrit and theanalyte may include glucose; the physical characteristic may include atleast one of viscosity, hematocrit, temperature and density; thedirecting may include driving first and second alternating signal atdifferent respective frequencies in which a first frequency is lowerthan the second frequency; the first frequency may be at least one orderof magnitude lower than the second frequency; the first frequency mayinclude any frequency in the range of about 10 kHz to about 250 kHz, orabout 10 kHz to about 90 kHz; and/or the specified sampling time may becalculated using an equation of the form:SpecifiedSamplingTime=x ₁ H ^(x) ² +x ₃where

“SpecifiedSamplingTime” is designated as a time point from the start ofthe test sequence at which to sample the output signal (e.g. outputsignal) of the test strip,

H represents, or is physical characteristic of the sample;

x₁ is about 4.3e5, or is equal to 4.3e5, or is equal to 4.3e5+/−10%, 5%or 1% of the numerical value provided hereof;

x₂ is about −3.9, or is equal to −3.9, or is equal to −3.9+/−10%, 5% or1% of the numerical value provided hereof; and

x₃ is about 4.8, or is equal to 4.8, or is equal to 4.8+/−10%, 5% or 1%of the numerical value provided herein. The term “e”, as used throughoutthis disclosure, refers to scientific notation. Accordingly, 4.3e5 is4.3×10⁵ or 430,000.

Additionally, the calculating of the analyte concentration may includeutilizing an equation of the form:

$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{Slope} \right\rbrack$where

G₀ represents, or is, an analyte concentration

I_(E) represents, or is, a signal (value or measurement; proportional toanalyte concentration) measured at the SpecifiedSamplingTime, which maybe the total signal measured at the SpecifiedSamplingTime;

Slope represents, or is, the value obtained from calibration testing ofa batch of test strips of which this particular strip comes from; and

Intercept represents, or is, the value obtained from calibration testingof a batch of test strips of which this particular strip comes from.

In addition to the above, applicants have provided various embodimentsof a technique to allow for improved glucose measurement using twosomewhat related relationships: (a) a relationship between sampling timepoint and hematocrit to derive or calculate a specified sampling time atwhich a measurement of the output from the biosensor is to be taken; and(b) a relationship between batch slope and physical characteristic(e.g., hematocrit) that allows derivation of a new batch slope. Bothrelationships are utilized to determine a more accurate analyteconcentration (i.e., based on the specified sampling time and thederived batch slope). This new technique does not rely on correction(s)or modification(s) to be made to an analyte measurement, therebyreducing test time while at the same time improving accuracy.

In an eleventh aspect of applicants' disclosure, a method of determiningan analyte concentration from a fluid sample (which may be aphysiological sample) with a biosensor is provided. The biosensor has atleast two electrodes and a reagent disposed on at least one electrode ofthe electrodes. The method can be achieved by: depositing a fluid sample(which may be a physiological sample) on the at least two electrodes tostart an analyte test sequence; applying a first signal to the sample toderive a physical characteristic of the sample; obtaining a physicalcharacteristic of the sample; specifying a sampling time based on thephysical characteristic from the obtaining step; deriving a batch slopefor the biosensor based on the physical characteristic from theobtaining step; driving a second signal to the sample; and measuring anoutput signal at the specified sampling time from at least one electrodeof the at least two electrodes; and calculating an analyte concentrationbased on the measured output signal at the specified sampling time andthe derived batch slope.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: theapplying of the first signal and the driving of the second signal may bein sequential order; the applying of the first signal may overlap withthe driving of the second signal; the applying of the first signal mayinclude directing an alternating signal to the sample so that a physicalcharacteristic of the sample is determined from an output of thealternating signal; the applying of the first signal may includedirecting an optical signal to the sample so that a physicalcharacteristic of the sample is determined from an output of the opticalsignal; the physical characteristic may include hematocrit and theanalyte may include glucose; the physical characteristic may include atleast one of viscosity, hematocrit, temperature and density of thesample; the directing may include driving first and second alternatingsignal at different respective frequencies in which a first frequency islower than the second frequency; the first frequency may be at least oneorder of magnitude lower than the second frequency; the first frequencymay include any frequency in the range of about 10 kHz to about 250 kHz,or about 10 kHz to about 90 kHz; and/or the specified sampling time maybe calculated using an equation of the form:SpecifiedSamplingTime=x ₁ H ^(x) ² +x ₃where

“SpecifiedSamplingTime” is designated as a time point from the start ofthe test sequence at which to sample the output signal of the teststrip,

H represents physical characteristic of the sample;

x₁ is about 4.3e5, or is equal to 4.3e5, or is equal to 4.3e5+/−10%, 5%or 1% of the numerical value provided hereof;

x₂ is about −3.9, or is equal to −3.9, or is equal to −3.9+/−10%, 5% or1% of the numerical value provided hereof; and

x₃ is about 4.8, or is equal to 4.8, or is equal to 4.8+/−10%, 5% or 1%of the numerical value provided herein. The term “e”, as used throughoutthis disclosure, refers to scientific notation. Accordingly, 4.3e5 is4.3×10⁵ or 430,000.

Additionally, for these aspects noted above, the derived slope may bedetermined from an equation of the form:NewSlope=aH ² +bH+cwhere

H is a measured or estimated physical characteristic (e.g., hematocrit);

a represents about 1.4e-6, or is equal to 1.35e-6, or is equal to1.4e-6+/−10%, 5% or 1% of the numerical value provided hereof;

b represents about −3.8e-4, or is equal to −3.79e-4, or is equal to−3.8e-4+/−10%, 5% or 1% of the numerical value provided hereof;

c represents about 3.6e-2, or is equal to 3.56e-2, or is equal to−3.6e-2+/−10%, 5% or 1% of the numerical value provided herein. The term“e”, as used throughout this disclosure, refers to scientific notation.Accordingly, 1.4e-6 is 1.4×10⁻⁶ or 0.0000014.

Furthermore, the calculating of the analyte concentration is computedwith an equation of the form:

$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{{New}{Slope}} \right\rbrack$where

G₀ represents, or is, an analyte concentration

I_(E) represents, or is, a signal (proportional to analyteconcentration) measured at the SpecifiedSamplingTime, which may be thetotal signal measured at the SpecifiedSamplingTime;

NewSlope represents, or is, the value derived from the measured orestimated physical characteristic; and

Intercept represents, or is, the value obtained from calibration testingof a batch of test strips of which this particular strip comes from.

In a twelfth aspect of applicants' disclosure, an analyte measurementsystem is provided that includes a test strip and an analyte meter. Thetest strip includes a substrate and a plurality of electrodes connectedto respective electrode connectors. The analyte meter includes ahousing, a test strip port connector configured to connect to therespective electrode connectors of the test strip, and a microprocessorin electrical communication with the test strip port connector to applyelectrical signals or sense electrical signals from the plurality ofelectrodes during a test sequence. The microprocessor is configured to,during the test sequence: (a) apply a first signal to the plurality ofelectrodes so that a specified sampling time and a batch slopedetermined from a physical characteristic of a physiological fluidsample are derived, (b) apply a second signal to the plurality ofelectrodes, and (c) measure a signal output from one of the plurality ofelectrodes at the specified sampling time so that an analyteconcentration is determined based on the measured signal at thespecified time point and the batch slope.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: theplurality of electrodes may include at least two electrodes to measurethe physical characteristic and at least two other electrodes to measurethe analyte concentration; the at least two electrodes and the at leasttwo other electrodes may be disposed in the same chamber provided on thesubstrate; the at least two electrodes and the at least two otherelectrodes may be disposed in different chambers provided on thesubstrate; the at least two electrodes may comprise two electrodes tomeasure the physical characteristic and the analyte concentration; theplurality of electrodes may comprise two electrodes to measure thephysical characteristic and the analyte concentration; all of theelectrodes may be disposed on the same plane defined by the substrate; areagent may be disposed proximate the at least two other electrodes andno reagent disposed on the at least two electrodes; and/or the specifiedsampling time may be calculated using an equation of the form:SpecifiedSamplingTime=x ₁ H ^(x) ² +x ₃where

“SpecifiedSamplingTime” is designated as a time point from the start ofthe test sequence at which to sample the output signal of the teststrip,

H represents, or is, a physical characteristic of the sample;

x₁ is about 4.3e5, or is equal to 4.3e5, or is equal to 4.3e5+/−10%, 5%or 1% of the numerical value provided hereof;

x₂ is about −3.9, or is equal to −3.9, or is equal to −3.9+/−10%, 5% or1% of the numerical value provided hereof; and

x₃ is about 4.8, or is equal to 4.8, or is equal to 4.8+/−10%, 5% or 1%of the numerical value provided herein. The term “e”, as used throughoutthis disclosure, refers to scientific notation. Accordingly, 4.3e5 is4.3×10⁵ or 430,000.

In these aspects previously noted above, the derived slope is determinedfrom an equation of the form:NewSlope=aH ² +bH+cwhere

H is measured or estimated physical characteristic (e.g., hematocrit);

a represents about 1.4e-6, or is equal to 1.35e-6, or is equal to1.4e-6+/−10%, 5% or 1% of the numerical value provided hereof;

b represents about −3.8e-4, or is equal to −3.79e-4, or is equal to−3.8e-4+/−10%, 5% or 1% of the numerical value provided hereof;

c represents about 3.6e-2, or is equal to 3.56e-2, or is equal to−3.6e-2+/−10%, 5% or 1% of the numerical value provided herein. The term“e”, as used throughout this disclosure, refers to scientific notation.Accordingly, 1.4e-6 is 1.4×10⁻⁶ or 0.0000014.

Furthermore, for these aspects, the calculating of the analyteconcentration is computed with an equation of the form:

$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{NewSlope} \right\rbrack$where

G₀ represents, or is, an analyte concentration

I_(E) represents, or is, a signal (proportional to analyteconcentration) measured at the SpecifiedSamplingTime, which may be thetotal signal measured at the SpecifiedSamplingTime;

NewSlope represents, or is, the value derived from the measured orestimated physical characteristic; and

Intercept represents, or is, the value obtained from calibration testingof a batch of test strips of which this particular strip comes from.

In a thirteenth aspect of applicants' disclosure, an analyte measurementsystem is provided that includes a test strip and an analyte meter. Thetest strip includes a substrate and a plurality of electrodes connectedto respective electrode connectors. The analyte meter includes ahousing, a test strip port connector configured to connect to therespective electrode connectors of the test strip, and a microprocessorin electrical communication with the test strip port connector to applyelectrical signals or sense electrical signals from the plurality ofelectrodes during a test sequence. The microprocessor is configured to,during a test sequence: (a) apply a first signal to the plurality ofelectrodes so that a specified sampling time and a batch slope of thetest strip determined from a physical characteristic of a physiologicalfluid sample are derived, (b) apply a second signal to the plurality ofelectrodes, and (c) measure a signal output from one of the plurality ofelectrodes at the specified sampling time point so that an analyteconcentration of the sample is determined based on the specifiedsampling time and batch slope within about 10 seconds of a start of thetest sequence.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: theplurality of electrodes may include at least two electrodes to measurethe physical characteristic and at least two other electrodes to measurethe analyte concentration; the at least two electrodes and the at leasttwo other electrodes may be disposed in the same chamber provided on thesubstrate; the at least two electrodes and the at least two otherelectrodes may be disposed in different chambers provided on thesubstrate; the at least two electrodes may comprise two electrodes tomeasure the physical characteristic and the analyte concentration; theplurality of electrodes may comprise two electrodes to measure thephysical characteristic and the analyte concentration; all of theelectrodes may be disposed on the same plane defined by the substrate; areagent may be disposed proximate the at least two other electrodes andno reagent disposed on the at least two electrodes; the specifiedsampling time may be calculated using an equation of the form:SpecifiedSamplingTime=x ₁ H ^(x) ² +x ₃where

“SpecifiedSamplingTime” is designated as a time point from the start ofthe test sequence at which to sample the output signal of the teststrip,

H represents, or is, a physical characteristic of the sample;

x₁ is about 4.3e5, or is equal to 4.3e5, or is equal to 4.3e5+/−10%, 5%or 1% of the numerical value provided hereof;

x₂ is about −3.9, or is equal to −3.9, or is equal to −3.9+/−10%, 5% or1% of the numerical value provided hereof; and

x₃ is about 4.8, or is equal to 4.8, or is equal to 4.8+/−10%, 5% or 1%of the numerical value provided herein. The term “e”, as used throughoutthis disclosure, refers to scientific notation. Accordingly, 4.3e5 is4.3×10⁵ or 430,000.

For these aspects previously noted, the derived slope may be determinedfrom an equation of the form:NewSlope=aH ² +bH+cwhere

NewSlope represents the derived slope;

H is measured or estimated physical characteristic (e.g., hematocrit);

a represents, or is, about 1.4e-6, or is equal to 1.35e-6, or is equalto 1.4e-6+/−10%, 5% or 1% of the numerical value provided hereof;

b represents, or is, about −3.8e-4, or is equal to −3.79e-4, or is equalto −3.8e-4+/−10%, 5% or 1% of the numerical value provided hereof;

c represents, or is, about 3.6e-2, or is equal to 3.56e-2, or is equalto −3.6e-2+/−10%, 5% or 1% of the numerical value provided hereof. Theterm “e”, as used throughout this disclosure, refers to scientificnotation. Accordingly, 1.4e-6 is 1.4×10⁻⁶ or 0.0000014.

Furthermore, for these aspects, the calculating of the analyteconcentration is computed with an equation of the form:

$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{NewSlope} \right\rbrack$where

G₀ represents, or is, an analyte concentration

I_(E) represents, or is, a signal (proportional to analyteconcentration) measured at the SpecifiedSamplingTime, which may be thetotal signal measured at the SpecifiedSamplingTime;

NewSlope represents, or is, the value derived from the measured orestimated physical characteristic; and

Intercept represents, or is, the value obtained from calibration testingof a batch of test strips of which this particular strip comes from.

In a fourteenth aspect of applicants' disclosure, a method ofdemonstrating increased accuracy of a test strip is provided. The methodcan be achieved by: providing a batch of test strips; introducing areferential sample containing a referential concentration of an analyteto each of the batch of test strips to initiate a test sequence;reacting the analyte to cause a physical transformation of the analyteproximate the two electrodes (which may be between the two electrodes);determining a physical characteristic of the referential sample;deriving a batch slope of the batch of test strips based on thedetermined physical characteristic; sampling an electrical output of thereferential sample at a specified time point during the test sequencedefined by the measured or estimated physical characteristic; andcalculating an analyte concentration based on the specified time pointand the derived batch slope to provide for a final analyte concentrationvalue for each of the batch of test strips such that at least 95% of thefinal analyte concentration values of the batch of test strips arewithin ±15% of the referential analyte concentration.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: thereacting may include driving a second signal to the sample and thedetermining may include applying a first signal to the sample to derivea physical characteristic of the sample, and the applying of the firstsignal and the driving of the second signal may be in sequential order;the applying of the first signal may overlap with the driving of thesecond signal; the reacting may comprise driving a second signal to thesample and the determining may comprise applying a first signal to thesample to derive a physical characteristic of the sample, and theapplying of the first signal may overlap with the driving of the secondsignal; the applying of the first signal may include directing analternating signal to the sample so that a physical characteristic ofthe sample is determined from an output of the alternating signal; theapplying of the first signal may include directing an optical signal tothe sample so that a physical characteristic of the sample is determinedfrom an output of the optical signal; the physical characteristic mayinclude hematocrit and the analyte may include glucose; the physicalcharacteristic may include at least one of viscosity, hematocrit,temperature and density; the directing may include driving first andsecond alternating signal at different respective frequencies in which afirst frequency is lower than the second frequency; the first frequencymay be at least one order of magnitude lower than the second frequency;the first frequency may include any frequency in the range of about 10kHz to about 250 kHz, or about 10 kHz to about 90 kHz; and/or thespecified sampling time may be calculated using an equation of the form:SpecifiedSamplingTime=x ₁ H ^(x) ² +x ₃where

“SpecifiedSamplingTime” is designated as a time point from the start ofthe test sequence at which to sample the output signal of the teststrip,

H represents, or is, a physical characteristic of the sample;

x₁ is about 4.3e5, or is equal to 4.3e5, or is equal to 4.3e5+/−10%, 5%or 1% of the numerical value provided hereof;

x₂ is about −3.9, or is equal to −3.9, or is equal to −3.9+/−10%, 5% or1% of the numerical value provided hereof and

x₃ is about 4.8, or is equal to 4.8, or is equal to 4.8+/−10%, 5% or 1%of the numerical value provided hereof. The term “e”, as used throughoutthis disclosure, refers to scientific notation. Accordingly, 4.3e5 is4.3×10⁵ or 430,000.

Additionally, for these previously disclosed aspects, the derived slopemay be determined from an equation of the form:NewSlope=aH ² +bH+cwhere

H is measured or estimated physical characteristic (e.g., hematocrit);

a represents, or is, about 1.4e-6, or is equal to 1.35e-6, or is equalto 1.4e-6+/−10%, 5% or 1% of the numerical value provided hereof

b represents, or is, about −3.8e-4, or is equal to −3.79e-4, or is equalto −3.8e-4+/−10%, 5% or 1% of the numerical value provided hereof

c represents, or is, about 3.6e-2, or is equal to 3.56e-2, or is equalto −3.6e-2+/−10%, 5% or 1% of the numerical value provided hereof. Theterm “e”, as used throughout this disclosure, refers to scientificnotation. Accordingly, 1.4e-6 is 1.4×10⁻⁶ or 0.0000014.

Furthermore, for these aspects, the calculating of the analyteconcentration is computed with an equation of the form:

$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{NewSlope} \right\rbrack$where

G₀ represents, or is, an analyte concentration

I_(E) represents, or is, a signal (proportional to analyteconcentration) or signals measured at the SpecifiedSamplingTime, whichmay be the total signal measured at the SpecifiedSamplingTime;

NewSlope represents, or is, the value derived from the measured orestimated physical characteristic; and

Intercept represents, or is, the value obtained from calibration testingof a batch of test strips of which this particular strip comes from.

In a fifteenth aspect of applicants' disclosure, a method of determiningan analyte concentration from a fluid sample (which may be aphysiological sample). The method can be achieved by: depositing a fluidsample (which may be a physiological sample) on a biosensor having areagent deposited thereon; applying signals to the sample and thereagent to transform the analyte into a different material; obtaining aphysical characteristic of the sample; specifying a time point forsampling of signal output based on the physical characteristic from theobtaining step; deriving a batch slope of the biosensor; measuringsignal output at the specified sampling time; and determining an analyteconcentration based on the measured signal output of the sample at thespecified sampling time and the derived batch slope.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: theobtaining may include driving a second signal to the sample to derive aphysical characteristic of the sample; the applying may include applyinga first signal to the sample to derive a physical characteristic of thesample, and the applying of the first signal and the driving of thesecond signal may be in sequential order; the applying of the firstsignal may overlap with the driving of the second signal; the applyingmay comprise applying a first signal to the sample to derive a physicalcharacteristic of the sample, and the applying of the first signal mayoverlap with the driving of the second signal; the applying of the firstsignal may include directing an alternating signal to the sample so thata physical characteristic of the sample is determined from an output ofthe alternating signal; the applying of the first signal may includedirecting an optical signal to the sample so that a physicalcharacteristic of the sample is determined from an output of the opticalsignal; the physical characteristic may include hematocrit and theanalyte may include glucose; the physical characteristic may include atleast one of viscosity, hematocrit, temperature and density; thedirecting may include driving first and second alternating signal atdifferent respective frequencies in which a first frequency is lowerthan the second frequency; the first frequency may be at least one orderof magnitude lower than the second frequency; the first frequency mayinclude any frequency in the range of about 10 kHz to about 250 kHz, orabout 10 kHz to about 90 kHz; and/or the specified sampling time may becalculated using an equation of the form:SpecifiedSamplingTime=x ₁ H ^(x) ² +x ₃where

“SpecifiedSamplingTime” is designated as a time point from the start ofthe test sequence at which to sample the output signal of the teststrip,

H represents, or is, a physical characteristic of the sample;

x₁ is about 4.3e5, or is equal to 4.3e5, or is equal to 4.3e5+/−10%, 5%or 1% of the numerical value provided hereof

x₂ is about −3.9, or is equal to −3.9, or is equal to −3.9+/−10%, 5% or1% of the numerical value provided hereof and

x₃ is about 4.8, or is equal to 4.8, or is equal to 4.8+/−10%, 5% or 1%of the numerical value provided hereof. The term “e”, as used throughoutthis disclosure, refers to scientific notation. Accordingly, 4.3e5 is4.3×10⁵ or 430,000.

For these aspects previously disclosed, the derived slope may bedetermined from an equation of the form:NewSlope=aH ² +bH+cwhere

H is measured or estimated physical characteristic (e.g., hematocrit);

a represents, or is, about 1.4e-6, or is equal to 1.35e-6, or is equalto 1.4e-6+/−10%, 5% or 1% of the numerical value provided hereof

b represents, or is, about −3.8e-4, or is equal to −3.79e-4, or is equalto −3.8e-4+/−10%, 5% or 1% of the numerical value provided hereof

c represents, or is, about 3.6e-2, or is equal to 3.56e-2, or is equalto −3.6e-2+/−10%, 5% or 1% of the numerical value provided hereof. Theterm “e”, as used throughout this disclosure, refers to scientificnotation. Accordingly, 1.4e-6 is 1.4×10⁻⁶ or 0.0000014.

Furthermore, the calculating of the analyte concentration is computedwith an equation of the form:

$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{NewSlope} \right\rbrack$where

G₀ represents, or is, an analyte concentration

I_(E) represents, or is, a signal (proportional to analyteconcentration) measured at the SpecifiedSamplingTime, which may be thetotal signal measured at the SpecifiedSamplingTime;

NewSlope represents, or is, the value derived from the measured orestimated physical characteristic; and

Intercept represents, or is, the value obtained from calibration testingof a batch of test strips of which this particular strip comes from.

In a sixteenth aspect of applicants' disclosure, a method of determiningan analyte concentration from a fluid sample (which may be aphysiological sample) with a biosensor is provided. The biosensor has atleast two electrodes and a reagent disposed on at least one of theelectrodes. The method can be achieved by: depositing a fluid sample(which may be a physiological sample) on the at least two electrodes tostart an analyte test sequence; applying a first signal to the sample tomeasure a physical characteristic of the sample; driving a second signalto the sample to cause an enzymatic reaction of the analyte and thereagent; estimating an analyte concentration based on a predeterminedsampling time point from the start of the test sequence; selecting asampling time point from a look-up table that includes a matrix in whichdifferent qualitative categories of the estimated analyte are set forthin the leftmost column of the matrix and different qualitativecategories of the measured or estimated physical characteristic are setforth in the topmost row of the matrix and the sampling times areprovided in the remaining cells of the matrix; measuring signal outputfrom the sample at the selected sampling time point from the look-uptable; calculating an analyte concentration from measured output signalsampled at said selected sampling time point in accordance with anequation of the form:

$G_{0} = \left\lbrack \frac{I_{T} - {Intercept}}{Slope} \right\rbrack$where

G₀ represents, or is, an analyte concentration;

I_(T) represents, or is, a signal (proportional to analyteconcentration) measured at a specified sampling time T, which may be thetotal signal measured at the specified sampling time T;

Slope represents, or is the value obtained from calibration testing of abatch of test strips of which this particular strip comes from; and

Intercept represents, or is, the value obtained from calibration testingof a batch of test strips of which this particular strip comes from.

In a seventeenth aspect of applicants' disclosure, a method ofdetermining an analyte concentration from a fluid sample (which may be aphysiological sample) with a biosensor is provided. The biosensor has atleast two electrodes and a reagent disposed on at least one of theelectrodes. The method can be achieved by: depositing a fluid sample(which may be a physiological sample) on the at least two electrodes tostart an analyte test sequence; applying a first signal to the sample tomeasure or estimate a physical characteristic of the sample; driving asecond signal to the sample to cause an enzymatic reaction of theanalyte and the reagent; estimating an analyte concentration based on apredetermined sampling time point from the start of the test sequence;selecting a sampling time point from a look-up table that includes amatrix in which different qualitative categories of the estimatedanalyte are set forth in the leftmost column of the matrix and differentqualitative categories of the measured or estimated physicalcharacteristic are set forth in the topmost row of the matrix and thesampling times are provided in the remaining cells of the matrix;measuring signal output from the sample at the selected sampling timepoint; and calculating an analyte concentration from measured outputsignal sampled at said selected sampling time point.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: theapplying of the first signal and the driving of the second signal may besequential; the applying of the first signal may overlap with thedriving of the second signal; the applying of the first signal maycomprise directing an alternating signal to the sample so that aphysical characteristic of the sample is determined from an output ofthe alternating signal; the physical characteristic may comprisehematocrit and the analyte may comprise glucose; the physicalcharacteristic may comprise at least one of viscosity, hematocrit,temperature and density; the directing may comprise driving first andsecond alternating signals at different respective frequencies in whicha first frequency is lower than the second frequency; the firstfrequency may be at least one order of magnitude lower than the secondfrequency; the first frequency may comprise any frequency in the rangeof about 10 kHz to about 250 kHz, or about 10 kHz to about 90 kHz; themeasuring may comprise sampling the signal output continuously at thestart of the test sequence until at least about 10 seconds after thestart; which may further include the step of estimating an analyteconcentration based on a measurement of the output signal at apredetermined time; the estimating may comprise comparing the estimatedanalyte concentration and the measured or estimated physicalcharacteristic against a look-up table having different respectiveranges of analyte concentration and physical characteristic of thesample indexed against different sample measurement times so that thepoint in time for measurement of the output from the sample of thesecond signal is obtained for the calculating step; and/or thecalculating step may comprise utilizing an equation of the form:

$G_{0} = \left\lbrack \frac{I_{T} - {Intercept}}{Slope} \right\rbrack$where

G₀ represents, or is, an analyte concentration;

I_(T) represents, or is, a signal (proportional to analyteconcentration) measured at a specified sampling time T, which may be thetotal signal measured at the specified sampling time T;

Slope represents, or is, the value obtained from calibration testing ofa batch of test strips of which this particular strip comes from; and

Intercept represents, or is, the value obtained from calibration testingof a batch of test strips of which this particular strip comes from.

In an eighteenth aspect of applicants' disclosure, an analytemeasurement system is provided that includes a test strip and an analytemeter. The test strip includes a substrate and a plurality of electrodesconnected to respective electrode connectors. The analyte meter includesa housing, a test strip port connector configured to connect to therespective electrode connectors of the test strip, and a microprocessorin electrical communication with the test strip port connector to applyelectrical signals or sense electrical signals from the plurality ofelectrodes. The microprocessor is configured to: (a) apply a firstsignal to the plurality of electrodes so that a physical characteristicof a physiological fluid sample is determined; (b) estimate an analyteconcentration based on a predetermined sampling time point during a testsequence; and (c) apply a second signal to the plurality of electrodesat a sampling time point during the test sequence dictated by thedetermined physical characteristic so that an analyte concentration iscalculated from the second signal.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: theplurality of electrodes may comprise at least two electrodes to measurethe physical characteristic and at least two other electrodes to measurethe analyte concentration; the at least two electrodes and the at leasttwo other electrodes may be disposed in the same chamber provided on thesubstrate; the at least two electrodes and the at least two otherelectrodes may be disposed in different chambers provided on thesubstrate; all of the electrodes may be disposed on the same planedefined by the substrate; and/or a reagent may be disposed proximate theat least two other electrodes and no reagent disposed on the at leasttwo electrodes.

In a nineteenth aspect of applicants' disclosure, an analyte measurementsystem is provided that includes a test strip and an analyte meter. Thetest strip includes a substrate and a plurality of electrodes connectedto respective electrode connectors. The analyte meter includes ahousing, a test strip port connector configured to connect to therespective electrode connectors of the test strip, and a microprocessorin electrical communication with the test strip port connector to applyelectrical signals or sense electrical signals from the plurality ofelectrodes. The microprocessor is configured to: (a) apply a firstsignal to the plurality of electrodes so that a physical characteristicof a physiological fluid sample is determined during a test sequence;(b) estimate an analyte concentration based on a predetermined samplingtime point during a test sequence; and (c) apply a second signal to theplurality of electrodes at a sampling time point during the testsequence dictated by the determined physical characteristic so that sothat an analyte concentration is determined from the second signalwithin about 10 seconds of a start of the test sequence.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: theplurality of electrodes may comprise at least two electrodes to measurethe physical characteristic and at least two other electrodes to measurethe analyte concentration; the at least two electrodes and the at leasttwo other electrodes may be disposed in the same chamber provided on thesubstrate; the at least two electrodes and the at least two otherelectrodes may be disposed in different chambers provided on thesubstrate; all of the electrodes may be disposed on the same planedefined by the substrate; and/or a reagent may be disposed proximate onthe at least two other electrodes and no reagent disposed on the atleast two electrodes.

In a twentieth aspect of applicants' disclosure, a method ofdemonstrating increased accuracy of a test strip is provided. The methodcan be achieved by: providing a batch of test strips; introducing areferential sample containing a referential concentration of an analyteto each test strip of the batch of test strips to start a test sequence;reacting the analyte with reagent disposed on each of the test strips tocause a physical transformation of the analyte proximate the twoelectrodes (which may be between the two electrodes); estimating ananalyte concentration based on measured signal output of the sample at apredetermined time point from the start of the test sequence;determining a physical characteristic of the referential sample;sampling an electrical output of the referential sample at a dictatedtime point during the test sequence defined by the measured or estimatedphysical characteristic and the estimated analyte concentration;calculating an analyte concentration based on the dictated time point toprovide for a final analyte concentration value for each test strip ofthe batch of test strips such that at least 95% of the final analyteconcentration values of the batch of test strips are within ±10% of thereferential analyte concentration for a range of hematocrit of thesample from about 30% to about 55%.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: thedetermining may comprise applying a first signal to the sample tomeasure a physical characteristic of the sample; the reacting maycomprise driving a second signal to the sample; the applying of thefirst signal and the driving of the second signal may be sequential; theapplying of the first signal may overlap with the driving of the secondsignal; the applying of the first signal may comprise directing analternating signal to the sample so that a physical characteristic ofthe sample is determined from an output of the alternating signal; theapplying of the first signal may comprise directing an electromagneticsignal to the sample so that a physical characteristic of the sample isdetermined from an output of the electromagnetic signal; the physicalcharacteristic may comprise hematocrit and the analyte may compriseglucose; the physical characteristic may comprise at least one ofviscosity, hematocrit, temperature and density; the directing maycomprise driving first and second alternating signals at differentrespective frequencies in which a first frequency is lower than thesecond frequency; the first frequency may be at least one order ofmagnitude lower than the second frequency; the first frequency maycomprise any frequency in the range of about 10 kHz to about 250 kHz, orabout 10 kHz to about 90 kHz; and/or the measuring may comprise samplingthe signal output continuously at the start of the test sequence untilat least about 10 seconds after the start; which may further include thestep of estimating an analyte concentration based on a measurement ofthe output signal at a predetermined time; the estimating may comprisecomparing the estimated analyte concentration and the measured orestimated physical characteristic against a look-up table havingdifferent respective ranges of analyte concentration and physicalcharacteristic of the sample indexed against different samplemeasurement times so that the point in time for measurement of theoutput from the sample of the second signal is obtained for thecalculating step.

In a twenty-first aspect of applicants' disclosure, a method ofdetermining an analyte concentration from a fluid sample (which may be aphysiological sample) is provided. The method can be achieved by:depositing a fluid sample (which may be a physiological sample) on abiosensor to start a test sequence; causing the analyte in the sample toundergo an enzymatic reaction; estimating an analyte concentration inthe sample; measuring at least one physical characteristic of thesample; defining a time point from the start of the test sequence tosample output signals of the biosensor based on the estimated analyteconcentration and at least one physical characteristic from themeasuring step; sampling output signals of the biosensor at the definedtime point; determining an analyte concentration from sampled signals atthe defined time point.

And for these aspects, the following features may also be utilized invarious combinations with these previously disclosed aspects: themeasuring may comprise applying a first signal to the sample to measurea physical characteristic of the sample; the causing step may comprisedriving a second signal to the sample; the measuring may compriseevaluating an output signal from the at least two electrodes at a pointin time after the start of the test sequence, in which the point in timeis set as a function of at least the measured or estimated physicalcharacteristic; the determining step may comprise calculating an analyteconcentration from the measured output signal at said point in time; theapplying of the first signal and the driving of the second signal may besequential; the applying of the first signal may overlap with thedriving of the second signal; the applying of the first signal maycomprise directing an alternating signal to the sample so that aphysical characteristic of the sample is determined from an output ofthe alternating signal; which may further include the step of estimatingan analyte concentration based on a predetermined sampling time pointfrom the start of the test sequence; the defining may comprise selectinga defined time point based on both the measured or estimated physicalcharacteristic and the estimated analyte concentration; the physicalcharacteristic may comprise hematocrit and the analyte may compriseglucose; the physical characteristic may comprise at least one ofviscosity, hematocrit, temperature and density; the directing maycomprise driving first and second alternating signal at differentrespective frequencies in which a first frequency is lower than thesecond frequency; the first frequency may be at least one order ofmagnitude lower than the second frequency; the first frequency maycomprise any frequency in the range of about 10 kHz to about 250 kHz, orabout 10 kHz to about 90 kHz; the measuring may comprise sampling thesignal output continuously at the start of the test sequence until atleast about 10 seconds after the start; which may further include thestep of estimating an analyte concentration based on a measurement ofthe output signal at a predetermined time; and/or the estimating maycomprise comparing the estimated analyte concentration and the measuredor estimated physical characteristic against a look-up table havingdifferent respective ranges of analyte concentration and physicalcharacteristic of the sample indexed against different samplemeasurement times so that the point in time for measurement of theoutput from the sample of the second signal is obtained for thecalculating step.

In the sixteenth to the twenty-first aspects, the sampling time pointcould be selected from a look-up table that includes a matrix in whichdifferent qualitative categories of the estimated analyte are set forthin the leftmost column of the matrix and different qualitativecategories of the measured or estimated physical characteristic are setforth in the topmost row of the matrix and the sampling times areprovided in the remaining cells of the matrix. In any of the aboveaspects, the fluid sample may be blood. In any of the above aspects, thephysical characteristic may include at least one of viscosity,hematocrit, or density of the sample, or the physical characteristic maybe hematocrit, wherein, optionally, the hematocrit level is between 30%and 55%. In any of the above aspects, where H represents, or is, thephysical characteristic of the sample, it may be the measured, estimatedor determined hematocrit, or may be in the form of hematocrit. In any ofthe above aspects, the physical characteristic may be determined from ameasured characteristic, such as the impedance or phase angle of thesample. In any of the above aspects, the signal represented by I_(E)and/or I_(T) may be current.

In the aforementioned aspects of the disclosure, the steps ofdetermining, estimating, calculating, computing, deriving and/orutilizing (possibly in conjunction with an equation) may be performed byan electronic circuit or a processor. These steps may also beimplemented as executable instructions stored on a computer readablemedium; the instructions, when executed by a computer may perform thesteps of any one of the aforementioned methods.

In additional aspects of the disclosure, there are computer readablemedia, each medium comprising executable instructions, which, whenexecuted by a computer, perform the steps of any one of theaforementioned methods.

In additional aspects of the disclosure, there are devices, such as testmeters or analyte testing devices, each device or meter comprising anelectronic circuit or processor configured to perform the steps of anyone of the aforementioned methods.

These and other embodiments, features and advantages will becomeapparent to those skilled in the art when taken with reference to thefollowing more detailed description of the exemplary embodiments of theinvention in conjunction with the accompanying drawings that are firstbriefly described.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings, which are incorporated herein and constitutepart of this specification, illustrate presently preferred embodimentsof the invention, and, together with the general description given aboveand the detailed description given below, serve to explain features ofthe invention (wherein like numerals represent like elements), in which:

FIG. 1 illustrates an analyte measurement system.

FIG. 2A illustrates in simplified schematic form the components of themeter 200.

FIG. 2B illustrates in simplified schematic form a preferredimplementation of a variation of meter 200.

FIG. 3A(1) illustrates the test strip 100 of the system of FIG. 1 inwhich there are two physical characteristic sensing electrodes upstreamof the measurement electrodes.

FIG. 3A(2) illustrates a variation of the test strip of FIG. 3A(1) inwhich a shielding or grounding electrode is provided for proximate theentrance of the test chamber.

FIG. 3A(3) illustrates a variation of the test strip of FIG. 3A(2) inwhich a reagent area has been extended upstream to cover at least one ofthe physical characteristic sensing electrodes.

FIG. 3A(4) illustrates a variation of test strip 100 of FIGS. 3A(1),3A(2) and 3A(3) in which certain components of the test strip have beenintegrated together into a single unit.

FIG. 3B illustrates a variation of the test strip of FIG. 3A(1), 3A(2),or 3A(3) in which one physical characteristic sensing electrode isdisposed proximate the entrance and the other physical characteristicsensing electrode is at the terminal end of the test cell with themeasurement electrodes disposed between the pair of physicalcharacteristic sensing electrodes.

FIGS. 3C and 3D illustrate variations of FIG. 3A(1), 3A(2), or 3A(3) inwhich the physical characteristic sensing electrodes are disposed nextto each other at the terminal end of the test chamber with themeasurement electrodes upstream of the physical characteristic sensingelectrodes.

FIGS. 3E and 3F illustrates a physical characteristic sensing electrodesarrangement similar to that of FIG. 3A(1), 3A(2), or 3A(3) in which thepair of physical characteristic sensing electrodes are proximate theentrance of the test chamber.

FIG. 3G is a simplified, perspective, exploded view of an analyticaltest strip according to an embodiment of the present disclosure.

FIG. 3H is a simplified top view of the analytical test strip of FIG.3G.

FIG. 3I is a simplified cross-sectional side view of the analytical teststrip of FIG. 3H taken along line A-A of FIG. 3H.

FIG. 3J is a simplified cross-sectional end view of the analytical teststrip of FIG. 3H taken along line B-B of FIG. 3H.

FIG. 3K is a simplified, perspective exploded view of an analytical teststrip according to an embodiment of the present disclosure.

FIG. 3L is a simplified top view of the electrically-insulatingsubstrate and a portion of a first patterned conductor layer of ananalytical test strip of FIG. 3K.

FIG. 3M is a simplified top view of the first patterned spacer layer ofthe analytical test strip of FIG. 3K.

FIG. 3N is a simplified top view of the second patterned spacer layer ofthe analytical test strip of FIG. 3K.

FIG. 3O is a simplified cross-sectional side view of the analytical teststrip of FIG. 3K taken along line A-A of FIG. 2A.

FIG. 3P is a simplified, perspective exploded view of an analytical teststrip according to another embodiment of the present disclosure.

FIG. 3Q is a simplified top view of the electrically insulatingsubstrate and first patterned conductor layer of the analytical teststrip of FIG. 3P.

FIG. 3R is a simplified top view of a portion of a second patternedspacer layer and second patterned conductor layer of the analytical teststrip of FIG. 3P.

FIG. 3S is a simplified top view of a third patterned spacer layer ofthe analytical test strip of FIG. 3P.

FIG. 3T is a simplified cross-sectional side view of the analytical teststrip of FIG. 3P taken along line B-B of FIG. 3Q.

FIG. 4A illustrates a graph of time over applied potential to the teststrip of FIG. 1.

FIG. 4B illustrates a graph of time over output current from the teststrip of FIG. 1.

FIG. 5 illustrates an exemplary waveform applied to the test chamber anda waveform as measured from the test chamber to show a time delaybetween the waveforms.

FIG. 6A(1) illustrates a logic diagram of an exemplary method to achievea more accurate analyte determination.

FIG. 6A(2) illustrates one relationship between batch slope and physicalcharacteristic (e.g., hematocrit).

FIG. 6A(3) illustrates data from test measurements conducted with theexemplary technique herein such that the data show the bias of less than±15% for the hematocrit range of about 30% to about 55%.

FIG. 6B(1) illustrates a logic diagram of an exemplary method to achievea more accurate analyte determination.

FIG. 6B(2) illustrates one relationship between sampling time point andhematocrits.

FIG. 6B(3) illustrates data from test measurements conducted with theexemplary technique herein such that the data show the bias of less than±25% for the hematocrit range of about 30% to about 55%.

FIG. 6C(1) illustrates a logic diagram of an exemplary method to achievea more accurate analyte determination.

FIG. 6C(2) illustrates one relationship between sampling time point andhematocrits.

FIG. 6C(3) illustrates a relationship between slope and hematocrits.

FIG. 6C(4) illustrates data from test measurements conducted with theexemplary technique herein such that the data show the bias of less than±25% for the hematocrit range of about 30% to about 55%.

FIG. 6D(1) illustrates a logic diagram of an exemplary method to achievea more accurate analyte determination.

FIG. 6D(2) illustrates a signal output transient of the biosensor andthe range of time point utilized for determination of the analyte, aswell as the estimation of the analyte concentration.

FIG. 6D(3) illustrates data from test measurements conducted with theexemplary technique herein such that the data show the bias of less thanabout ±10% for the hematocrit range of about 30% to about 55%.

FIG. 7 illustrates a signal output transient of the biosensor and therange of time point utilized for determination of the analyte, as wellas the estimation of the analyte concentration.

FIG. 8 is a simplified depiction of a hand-held test meter according toan embodiment of the present disclosure.

FIG. 9 is a simplified block diagram of various blocks of the hand-heldtest meter of FIG. 8.

FIG. 10 is a simplified block diagram of a phase-shift-based hematocritmeasurement block as can be employed in embodiments according to thepresent disclosure.

FIG. 11 is a simplified annotated schematic diagram of a dual low passfilter sub-block as can be employed in embodiments of the presentdisclosure.

FIG. 12 is a simplified annotated schematic diagram of a transimpedanceamplifier (TIA) sub-block as can be employed in embodiments of thepresent invention.

FIG. 13 is a simplified annotated schematic block diagram depicting adual low pass filter sub-block, a calibration load sub-block, ananalytical test strip sample cell interface sub-block, a transimpedanceamplifier sub-block, an XOR phase shift measurement sub-block and aQuadratur DEMUX phase-shift measurement sub-block as can be employed ina phase-shift-based hematocrit measurement block of embodiments of thepresent disclosure.

FIG. 14 is a flow diagram depicting stages in a method for employing ahand-held test meter according to an embodiment of the presentdisclosure.

MODES OF CARRYING OUT THE INVENTION

The following detailed description should be read with reference to thedrawings, in which like elements in different drawings are identicallynumbered. The drawings, which are not necessarily to scale, depictselected embodiments and are not intended to limit the scope of theinvention. The detailed description illustrates by way of example, notby way of limitation, the principles of the invention. This descriptionwill clearly enable one skilled in the art to make and use theinvention, and describes several embodiments, adaptations, variations,alternatives and uses of the invention, including what is presentlybelieved to be the best mode of carrying out the invention.

As used herein, the terms “about” or “approximately” for any numericalvalues or ranges indicate a suitable dimensional tolerance that allowsthe part or collection of components to function for its intendedpurpose as described herein. More specifically, “about” or“approximately” may refer to the range of values±10% of the recitedvalue, e.g. “about 90%” may refer to the range of values from 81% to99%. In addition, as used herein, the terms “patient,” “host,” “user,”and “subject” refer to any human or animal subject and are not intendedto limit the systems or methods to human use, although use of thesubject invention in a human patient represents a preferred embodiment.As used herein, “oscillating signal” includes voltage signal(s) orcurrent signal(s) that, respectively, change polarity or alternatedirection of current or are multi-directional. Also used herein, thephrase “electrical signal” or “signal” is intended to include directcurrent signal, alternating signal or any signal within theelectromagnetic spectrum. The terms “processor”; “microprocessor”; or“microcontroller” are intended to have the same meaning and are intendedto be used interchangeably. Finally, the term “e” as used in each of themathematical relationships described herein is intended to refer toscientific or exponential notation; e.g., 4.3e5=4.3×10⁵=430,000.

FIG. 1 illustrates a test meter 200, for testing analyte (e.g., glucose)levels in the blood of an individual with a test strip produced by themethods and techniques illustrated and described herein. Test meter 200may include user interface inputs (206, 210, 214), which can be in theform of buttons, for entry of data, navigation of menus, and executionof commands. Data can include values representative of analyteconcentration, and/or information that are related to the everydaylifestyle of an individual. Information, which is related to theeveryday lifestyle, can include food intake, medication use, theoccurrence of health check-ups, general health condition and exerciselevels of an individual. Test meter 200 can also include a display 204that can be used to report measured glucose levels, and to facilitateentry of lifestyle related information.

Test meter 200 may include a first user interface input 206, a seconduser interface input 210, and a third user interface input 214. Userinterface inputs 206, 210, and 214 facilitate entry and analysis of datastored in the testing device, enabling a user to navigate through theuser interface displayed on display 204. User interface inputs 206, 210,and 214 include a first marking 208, a second marking 212, and a thirdmarking 216, which help in correlating user interface inputs tocharacters on display 204.

Test meter 200 can be turned on by inserting a test strip 100 (or itsvariants 400, 500, or 600) into a strip port connector 220, by pressingand briefly holding first user interface input 206, or by the detectionof data traffic across a data port 218. Test meter 200 can be switchedoff by removing test strip 100 (or its variants 400, 500, or 600),pressing and briefly holding first user interface input 206, navigatingto and selecting a meter off option from a main menu screen, or by notpressing any buttons for a predetermined time. Display 104 canoptionally include a backlight.

In one embodiment, test meter 200 can be configured to not receive acalibration input for example, from any external source, when switchingfrom a first test strip batch to a second test strip batch. Thus, in oneexemplary embodiment, the meter is configured to not receive acalibration input from external sources, such as a user interface (suchas inputs 206, 210, 214), an inserted test strip, a separate code key ora code strip, data port 218. Such a calibration input is not necessarywhen all of the test strip batches have a substantially uniformcalibration characteristic. The calibration input can be a set of valuesascribed to a particular test strip batch. For example, the calibrationinput can include a batch slope and a batch intercept value for aparticular test strip batch. The calibrations input, such as batch slopeand intercept values, may be preset within the meter as will bedescribed below.

Referring to FIG. 2A, an exemplary internal layout of test meter 200 isshown. Test meter 200 may include a processor 300, which in someembodiments described and illustrated herein is a 32-bit RISCmicrocontroller. In the preferred embodiments described and illustratedherein, processor 300 is preferably selected from the MSP 430 family ofultra-low power microcontrollers manufactured by Texas Instruments ofDallas, Tex. The processor can be bi-directionally connected via I/Oports 314 to a memory 302, which in some embodiments described andillustrated herein is an EEPROM. Also connected to processor 300 via I/Oports 214 are the data port 218, the user interface inputs 206, 210, and214, and a display driver 320. Data port 218 can be connected toprocessor 300, thereby enabling transfer of data between memory 302 andan external device, such as a personal computer. User interface inputs206, 210, and 214 are directly connected to processor 300. Processor 300controls display 204 via display driver 320. Memory 302 may bepre-loaded with calibration information, such as batch slope and batchintercept values, during production of test meter 200. This pre-loadedcalibration information can be accessed and used by processor 300 uponreceiving a suitable signal (such as current) from the strip via stripport connector 220 so as to calculate a corresponding analyte level(such as blood glucose concentration) using the signal and thecalibration information without receiving calibration input from anyexternal source.

In embodiments described and illustrated herein, test meter 200 mayinclude an Application Specific Integrated Circuit (ASIC) 304, so as toprovide electronic circuitry used in measurements of glucose level inblood that has been applied to a test strip 100 (or its variants 400,500, or 600) inserted into strip port connector 220. Analog voltages canpass to and from ASIC 304 by way of an analog interface 306. Analogsignals from analog interface 306 can be converted to digital signals byan A/D converter 316. Processor 300 further includes a core 308, a ROM310 (containing computer code), a RAM 312, and a clock 318. In oneembodiment, the processor 300 is configured (or programmed) to disableall of the user interface inputs except for a single input upon adisplay of an analyte value by the display unit such as, for example,during a time period after an analyte measurement. In an alternativeembodiment, the processor 300 is configured (or programmed) to ignoreany input from all of the user interface inputs except for a singleinput upon a display of an analyte value by the display unit. Detaileddescriptions and illustrations of the meter 200 are shown and describedin International Patent Application Publication No. WO2006070200, whichis hereby incorporated by reference into this application as if fullyset forth herein.

FIG. 3A(1) is an exemplary exploded perspective view of a test strip100, which may include seven layers disposed on a substrate 5. The sevenlayers disposed on substrate 5 can be a first conductive layer 50 (whichcan also be referred to as electrode layer 50), an insulation layer 16,two overlapping reagent layers 22 a and 22 b, an adhesive layer 60 whichincludes adhesive portions 24, 26, and 28, a hydrophilic layer 70, and atop layer 80 which forms a cover 94 for the test strip 100. Test strip100 may be manufactured in a series of steps where the conductive layer50, insulation layer 16, reagent layers 22, and adhesive layer 60 aresequentially deposited on substrate 5 using, for example, ascreen-printing process. Note that the electrodes 10, 12, and 14) aredisposed for contact with the reagent layer 22 a and 22 b whereas thephysical characteristic sensing electrodes 19 a and 20 a are spacedapart and not in contact with the reagent layer 22. Hydrophilic layer 70and top layer 80 can be disposed from a roll stock and laminated ontosubstrate 5 as either an integrated laminate or as separate layers. Teststrip 100 has a distal portion 3 and a proximal portion 4 as shown inFIG. 3A(1).

Test strip 100 may include a sample-receiving chamber 92 through which aphysiological fluid sample 95 may be drawn through or deposited (FIG.3A(2)). The physiological fluid sample discussed herein may be blood.Sample-receiving chamber 92 can include an inlet at a proximal end andan outlet at the side edges of test strip 100, as illustrated in FIG.3A(1). A fluid sample 95 can be applied to the inlet along axis L-L(FIG. 3A(2)) to fill a sample-receiving chamber 92 so that glucose canbe measured. The side edges of a first adhesive pad 24 and a secondadhesive pad 26 located adjacent to reagent layer 22 each define a wallof sample-receiving chamber 92, as illustrated in FIG. 3A(1). A bottomportion or “floor” of sample-receiving chamber 92 may include a portionof substrate 5, conductive layer 50, and insulation layer 16, asillustrated in FIG. 3A(1). A top portion or “roof” of sample-receivingchamber 92 may include distal hydrophilic portion 32, as illustrated inFIG. 3A(1). For test strip 100, as illustrated in FIG. 3A(1), substrate5 can be used as a foundation for helping support subsequently appliedlayers. Substrate 5 can be in the form of a polyester sheet such as apolyethylene tetraphthalate (PET) material (Hostaphan PET supplied byMitsubishi). Substrate 5 can be in a roll format, nominally 350 micronsthick by 370 millimeters wide and approximately 60 meters in length.

A conductive layer is required for forming electrodes that can be usedfor the electrochemical measurement of glucose. First conductive layer50 can be made from a carbon ink that is screen-printed onto substrate5. In a screen-printing process, carbon ink is loaded onto a screen andthen transferred through the screen using a squeegee. The printed carbonink can be dried using hot air at about 140° C. The carbon ink caninclude VAGH resin, carbon black, graphite (KS15), and one or moresolvents for the resin, carbon and graphite mixture. More particularly,the carbon ink may incorporate a ratio of carbon black:VAGH resin ofabout 2.90:1 and a ratio of graphite:carbon black of about 2.62:1 in thecarbon ink.

For test strip 100, as illustrated in FIG. 3A(1), first conductive layer50 may include a reference electrode 10, a first working electrode 12, asecond working electrode 14, third and fourth physical characteristicsensing electrodes 19 a and 19 b, a first contact pad 13, a secondcontact pad 15, a reference contact pad 11, a first working electrodetrack 8, a second working electrode track 9, a reference electrode track7, and a strip detection bar 17. The physical characteristic sensingelectrodes 19 a and 20 a are provided with respective electrode tracks19 b and 20 b. The conductive layer may be formed from carbon ink. Firstcontact pad 13, second contact pad 15, and reference contact pad 11 maybe adapted to electrically connect to a test meter. First workingelectrode track 8 provides an electrically continuous pathway from firstworking electrode 12 to first contact pad 13. Similarly, second workingelectrode track 9 provides an electrically continuous pathway fromsecond working electrode 14 to second contact pad 15. Similarly,reference electrode track 7 provides an electrically continuous pathwayfrom reference electrode 10 to reference contact pad 11. Strip detectionbar 17 is electrically connected to reference contact pad 11. Third andfourth electrode tracks 19 b and 20 b connect to the respectiveelectrodes 19 a and 20 a. A test meter can detect that test strip 100has been properly inserted by measuring a continuity between referencecontact pad 11 and strip detection bar 17, as illustrated in FIG. 3A(1).

Variations of the test strip 100 (FIG. 3A(1), 3A(2), 3A(3), or 3A(4))are shown in FIGS. 3B-3T. Briefly, with regard to variations of teststrip 100 (illustrated exemplarily in FIGS. 3A(2), 3A(2) and 3B through3T), these test strips include an enzymatic reagent layer disposed onthe working electrode, a patterned spacer layer disposed over the firstpatterned conductive layer and configured to define a sample chamberwithin the analytical test strip, and a second patterned conductivelayer disposed above the first patterned conductive layer. The secondpatterned conductive layer includes a first phase-shift measurementelectrode and a second phase-shift measurement electrode. Moreover, thefirst and second phase-shift measurement electrodes are disposed in thesample chamber and are configured to measure, along with the hand-heldtest meter, a phase shift of an electrical signal forced through abodily fluid sample introduced into the sample chamber during use of theanalytical test strip. Such phase-shift measurement electrodes are alsoreferred to herein as bodily fluid phase-shift measurement electrodes.Analytical test strips of various embodiments described herein arebelieved to be advantageous in that, for example, the first and secondphase-shift measurement electrodes are disposed above the working andreference electrodes, thus enabling a sample chamber of advantageouslylow volume. This is in contrast to a configuration wherein the first andsecond phase-shift measurement electrodes are disposed in a co-planarrelationship with the working and reference electrodes thus requiring alarger bodily fluid sample volume and sample chamber to enable thebodily fluid sample to cover the first and second phase-shiftmeasurement electrodes as well as the working and reference electrodes.

In the embodiment of FIG. 3A(2) which is a variation of the test stripof FIG. 3A(1), an additional electrode 10 a is provided as an extensionof any of the plurality of electrodes 19 a, 20 a, 14, 12, and 10. Itmust be noted that the built-in shielding or grounding electrode 10 a isused to reduce or eliminate any capacitance coupling between the fingeror body of the user and the characteristic measurement electrodes 19 aand 20 a. The grounding electrode 10 a allows for any capacitance to bedirected away from the sensing electrodes 19 a and 20 a. To do this, thegrounding electrode 10 a can be connected any one of the other fiveelectrodes or to its own separate contact pad (and track) for connectionto ground on the meter instead of one or more of contact pads 15, 17, 13via respective tracks 7, 8, and 9. In a preferred embodiment, thegrounding electrode 10 a is connected to one of the three electrodesthat has reagent 22 disposed thereon. In a most preferred embodiment,the grounding electrode 10 a is connected to electrode 10. Being thegrounding electrode, it is advantageous to connect the groundingelectrode to the reference electrode (10) so not to contribute anyadditional current to the working electrode measurements which may comefrom background interfering compounds in the sample. Further byconnecting the shield or grounding electrode 10 a to electrode 10 thisis believed to effectively increase the size of the counter electrode 10which can become limiting especially at high signals. In the embodimentof FIG. 3A(2), the reagent are arranged so that they are not in contactwith the measurement electrodes 19 a and 20 a. Alternatively, in theembodiment of FIG. 3A(3), the reagent 22 is arranged so that the reagent22 contacts at least one of the sensing electrodes 19 a and 20 a.

In alternate version of test strip 100, shown here in FIG. 3A(4), thetop layer 38, hydrophilic film layer 34 and spacer 29 have been combinedtogether to form an integrated assembly for mounting to the substrate 5with reagent layer 22′ disposed proximate insulation layer 16′.

In the embodiment of FIG. 3B, the analyte measurement electrodes 10, 12,and 14 are disposed in generally the same configuration as in FIG.3A(1), 3A(2), or 3A(3). The electrodes 19 a and 20 a to sense physicalcharacteristic (e.g., hematocrit) level, however, are disposed in aspaced apart configuration in which one electrode 19 a is proximate anentrance 92 a to the test chamber 92 and another electrode 20 a is atthe opposite end of the test chamber 92. Electrodes 10, 12, and 14 aredisposed to be in contact with a reagent layer 22.

In FIGS. 3C, 3D, 3E and 3F, the physical characteristic (e.g.,hematocrit) sensing electrodes 19 a and 20 a are disposed adjacent eachother and may be placed at the opposite end 92 b of the entrance 92 a tothe test chamber 92 (FIGS. 3C and 3D) or adjacent the entrance 92 a(FIGS. 3E and 3F). In all of these embodiments, the physicalcharacteristic sensing electrodes are spaced apart from the reagentlayer 22 so that these physical characteristic sensing electrodes arenot impacted by the electrochemical reaction of the reagent in thepresence of a fluid sample (e.g., blood or interstitial fluid)containing glucose.

Referring to FIGS. 3G through 3J, electrochemical-based analytical teststrip 400 includes an electrically-insulating substrate layer 402, afirst patterned conductive layer 404 disposed on theelectrically-insulating substrate layer, an enzymatic reagent layer 406(for clarity depicted in FIG. 3G only), a patterned spacer layer 408, asecond patterned conductive layer 410 disposed above first patternedconductive layer 404, and an electrically-insulating top layer 412.Patterned spacer layer 408 is configured such that electrochemical-basedanalytical test strip 400 also includes a sample chamber 414 formedtherein with patterned spacer layer 408 defining outer walls of samplechamber 414.

First patterned conductive layer 404 includes three electrodes, acounter electrode 404 a (also referred to as a reference electrode), afirst working electrode 404 b and a second working electrode 404 c (seeFIG. 3G).

Second patterned conductive layer 410 includes a first phase-shiftmeasurement electrode 411 and a second phase shift measurement electrode413. Second patterned conductive layer 410 also includes a firstphase-shift probe contact 416 and a second phase-shift probe contact418.

During use of electrochemical-based analytical test strip 400 todetermine an analyte in a bodily fluid sample (e.g., blood glucoseconcentration in a whole physiological fluid sample), electrodes 404 a,404 b and 404 c are employed by an associated meter (not shown) tomonitor an electrochemical response of the electrochemical-basedanalytical test strip. The electrochemical response can be, for example,an electrochemical reaction induced current of interest. The magnitudeof such a current can then be correlated, taking into consideration thephysical characteristic (e.g., hematocrit) of the bodily fluid sample asdetermined by the bodily fluid sample's phase shift, with the amount ofanalyte present in the bodily fluid sample under investigation. Duringsuch use, a bodily fluid sample is applied to electrochemical-basedanalytical test strip 400 and, thereby, received in sample chamber 414.

Electrically-insulating substrate layer 402 can be any suitableelectrically-insulating substrate known to one skilled in the artincluding, for example, a nylon substrate, polycarbonate substrate, apolyimide substrate, a polyvinyl chloride substrate, a polyethylenesubstrate, a polypropylene substrate, a glycolated polyester (PETG)substrate, a polystyrene substrate, a silicon substrate, ceramicsubstrate, glass substrate or a polyester substrate (e.g., a 7millimeters thick polyester substrate). The electrically-insulatingsubstrate can have any suitable dimensions including, for example, awidth dimension of about 5 mm, a length dimension of about 27 mm and athickness dimension of about 0.5 mm.

First patterned conductive layer 404 can be formed of any suitableelectrically conductive material such as, for example, gold, palladium,carbon, silver, platinum, tin oxide, iridium, indium, or combinationsthereof (e.g., indium doped tin oxide). Moreover, any suitable techniqueor combination of techniques can be employed to form first patternedconductive layer 404 including, for example, sputtering, evaporation,electro-less plating, screen-printing, contact printing, laser ablationor gravure printing. A typical but non-limiting thickness for thepatterned conductive layer is in the range of 5 nanometers to 400nanometers.

As is known, conventional electrochemical-based analyte test stripsemploy a working electrode along with an associated counter/referenceelectrode and enzymatic reagent layer to facilitate an electrochemicalreaction with an analyte of interest and, thereby, determine thepresence and/or concentration of that analyte. For example, anelectrochemical-based analyte test strip for the determination ofglucose concentration in a fluid sample can employ an enzymatic reagentthat includes the enzyme glucose oxidase and the mediator ferricyanide(which is reduced to the mediator ferrocyanide during theelectrochemical reaction). Such conventional analyte test strips andenzymatic reagent layers are described in, for example, U.S. Pat. Nos.5,708,247; 5,951,836; 6,241,862; and 6,284,125; each of which is herebyincorporated by reference herein to this application. In this regard,the reagent layer employed in various embodiments provided herein caninclude any suitable sample-soluble enzymatic reagents, with theselection of enzymatic reagents being dependent on the analyte to bedetermined and the bodily fluid sample. For example, if glucose is to bedetermined in a fluid sample, enzymatic reagent layer 406 can includeglucose oxidase or glucose dehydrogenase along with other componentsnecessary for functional operation.

In general, enzymatic reagent layer 406 includes at least an enzyme anda mediator. Examples of suitable mediators include, for example,ruthenium, Hexaammine Ruthenium (III) Chloride, ferricyanide, ferrocene,ferrocene derivatives, osmium bipyridyl complexes, and quinonederivatives. Examples of suitable enzymes include glucose oxidase,glucose dehydrogenase (GDH) using a pyrroloquinoline quinone (PQQ)co-factor, GDH using a nicotinamide adenine dinucleotide (NAD)co-factor, and GDH using a flavin adenine dinucleotide (FAD) co-factor.Enzymatic reagent layer 406 can be applied during manufacturing usingany suitable technique including, for example, screen printing.

Applicants note that enzymatic reagent layer 406 may also containsuitable buffers (such as, for example, Tris HCl, Citraconate, Citrateand Phosphate), hydroxyethylcelulose [HEC], carboxymethylcellulose,ethycellulose and alginate, enzyme stabilizers and other additives asare known in the field.

Further details regarding the use of electrodes and enzymatic reagentlayers for the determination of the concentrations of analytes in abodily fluid sample, albeit in the absence of the phase-shiftmeasurement electrodes, analytical test strips and related methodsdescribed herein, are in U.S. Pat. No. 6,733,655, which is hereby fullyincorporated by reference herein to this application.

Patterned spacer layer 408 can be formed of any suitable materialincluding, for example, a 95 micrometers thick, double-sided pressuresensitive adhesive layer, a heat activated adhesive layer, or athermo-setting adhesive plastic layer. Patterned spacer layer 408 canhave, for example, a thickness in the range of from about 1 micron toabout 500 microns, preferably between about 10 microns and about 400microns, and more preferably between about 40 microns and about 200microns.

Second patterned conductive layer 410 can be formed of any suitableconductive material including, for example, copper, silver, palladium,gold and conductive carbon materials. Second patterned conductive layer410 can be, for example, disposed on a lower surface ofelectrically-insulating top layer 412 (as depicted in FIGS. 3G-3J) orembedded in the lower surface of electrically-insulating top layer 412.Second patterned conductive layer 410 can have any suitable thicknessincluding, for example, a thickness in the range of 20 microns to 400microns.

First phase-shift measurement electrode 411 and second phase shiftmeasurement electrode 413 of second patterned conductive layer 410 areseparated within sample chamber 414 by a gap (in the horizontaldirection of FIG. 3J) that is suitable for phase-shift measurement. Sucha gap can be, for example, in the range of 20 microns to 1,400 micronswith a typical gap being 500 microns. Moreover, the surface area offirst phase-shift measurement electrode 111 and second phase shiftmeasurement electrode 113 that is exposed to a bodily fluid samplewithin sample chamber 414 is typically 0.5 mm² but can range, forexample, from 0.1 mm² to 2.0 mm².

Electrochemical-based analytical test strip 400 can be manufactured, forexample, by the sequential aligned formation of first patternedconductive layer 404, enzymatic reagent layer 406, patterned spacerlayer 408, second patterned conductive layer 410 and electricallyinsulating top layer 412 onto electrically-insulating substrate layer402. Any suitable techniques known to one skilled in the art can be usedto accomplish such sequential aligned formation, including, for example,screen printing, photolithography, photogravure, chemical vapourdeposition, sputtering, tape lamination techniques and combinationsthereof.

Analytical test strips according to embodiments can be configured, forexample, for operable electrical connection (via, for example, first andsecond phase shift probe contacts 416 and 418) and use with theanalytical test strip sample cell interface of a hand-held test meter asdescribed in co-pending patent application Ser. No. 13/250,525, which ishereby incorporated by reference herein to this application with a copyprovided in the Appendix.

It has been determined that a relationship exists between the reactanceof a whole fluid sample and the physical characteristic (e.g.,hematocrit) of that sample. Electrical modeling of a bodily fluid sample(e.g., a whole blood sample) as parallel capacitive and resistivecomponents indicates that when an alternating current (AC) signal isforced through the bodily fluid sample, the phase shift of thealternating signal will be dependent on both the frequency of thealternating signal voltage and the physical characteristic (e.g.,hematocrit) of the sample. Therefore, the physical characteristic (e.g.,hematocrit) of a bodily fluid sample can be measured by, for example,driving alternating signals of a known frequency (or known frequencies)through the bodily fluid sample and detecting their phase shift. Thephase-shift measurement electrodes of analytical test strips of variousembodiments described herein are particularly suitable for use in suchphase-shift measurements since the first and second phase shiftmeasurement electrodes are in direct contact with a bodily fluid samplepresent in the sample chamber. Moreover, a bodily fluid sample physicalcharacteristic (e.g., hematocrit) ascertained from a phase shiftmeasurement(s) can be employed to compensate for the effect of physicalcharacteristic (e.g., hematocrit) during analyte determination.

Applicants note that for various embodiments of analytical test strips(e.g., an electrochemical-based analytical test strip) described herefor use with a hand-held test meter in the determination of an analyte(such as glucose) in a bodily fluid sample (for example, a whole bloodsample) may include an electrically insulating substrate, a firstpatterned conductor layer disposed on the electrically insulatingsubstrate and having a working electrode and a reference electrode. Theanalytical test strip may also include an enzymatic reagent layerdisposed on the working electrode, a first patterned spacer layerdisposed over the first patterned conductor layer and defining both afirst sample-receiving channel and an analyte determination samplechamber within the analytical test strip, and a second patterned spacerlayer disposed over the first patterned spacer layer and defining atleast a second sample-receiving channel. In addition, the analyticaltest strip further includes a bodily fluid phase-shift sample chamber influidic communication with the second sample-receiving channel.Moreover, the first sample-receiving channel and analyte determinationsample chamber of the analytical test strip are isolated from the secondsample-receiving channel and bodily fluid phase-shift sample chamber ofthe analytical test strip.

Analytical test strips of various embodiments described herein arebelieved by applicants to be beneficial in that, for example, theisolation (fluidic and electrical) between the analyte determinationsample chamber and the bodily fluid phase-shift sample chamber preventspotential interference between the determination of the analyte in thebodily fluid sample and a phase-shift measurement of the bodily fluid.Applicants note that certain advantages are obtained by having the firstsample-receiving channel and analyte determination chamber separatedfrom the second sample-receiving channel and bodily fluid phase-shiftsample chamber by portions of the first and/or second patterned spacerlayers that can be thinner, thus providing for an analytical test stripwith a small, yet mechanically stable, cross-section.

Referring to FIGS. 3K-3O, electrochemical-based analytical test strip500 includes an electrically-insulating substrate 502, a first patternedconductor layer 504 disposed on the electrically-insulating substratelayer, an enzymatic reagent layer 506 (for clarity depicted in FIG. 3Konly), a first patterned spacer layer 508, a second patterned spacerlayer 510, and a top cover 511. In the embodiment of FIG. 3K, firstpattered spacer layer 508 and second patterned spacer layer 510 aredepicted as bi-layer structures. However, the first and second patternedspacer layers employed in various embodiments provided herein can beunitary layers or any other suitably formed layer.

First patterned spacer layer 508 is configured such thatelectrochemical-based analytical test strip 500 also includes a firstsample-receiving channel 512 and an analyte determination sample chamber514. First patterned spacer layer 508 is also configured to define abodily fluid phase-shift sample chamber 516 and an analyte determinationsample chamber vent 518 (for clarity not depicted in FIG. 3K).

Second patterned spacer layer 510 is configured to define a secondsample-receiving channel 520 and a bodily fluid phase-shift chamber vent522 (for clarity not depicted in FIG. 3K).

First patterned conductor layer 504 includes a first phase-shiftmeasurement electrode 524, a second phase-shift measurement electrode526, two working electrodes 528 a and 528 b and a reference electrode530. For clarity, FIG. 3L depicts only first phase-shift measurementelectrode 524 and second phase-shift measurement electrode 526 and notthe entirety of first patterned conductor layer 504.

First sample-receiving channel 512 and analyte determination samplechamber 514 are isolated, both fluidically and electrically, from secondsample-receiving channel 520 and bodily fluid phase-shift sample chamber516 (see FIG. 3O in particular wherein the first and second patternedconductor layers are omitted for clarity). Moreover, in the embodimentof FIG. 3O, the bodily fluid phase-shift sample chamber is disposed in aside-by-side configuration with the analyte determination samplechamber.

During use of electrochemical-based analytical test strip 500 todetermine an analyte in a bodily fluid sample (e.g., blood glucoseconcentration in a whole blood sample), working and reference electrodesare employed by an associated meter (not shown) to monitor anelectrochemical response of the electrochemical-based analytical teststrip. The electrochemical response can be, for example, anelectrochemical reaction induced current of interest. The magnitude ofsuch a signal can then be correlated, taking into consideration thehaematocrit of the bodily fluid sample as determined by the bodily fluidsample's phase shift, with the amount of analyte present in the bodilyfluid sample under investigation. During such use, a bodily fluid sampleis applied to electrochemical-based analytical test strip 500 and,thereby, received in both analyte determination sample chamber 514 andbodily fluid phase-shift sample chamber 516.

Electrically-insulating substrate 502 can be any suitableelectrically-insulating substrate known to one skilled in the artincluding, for example, a nylon substrate, polycarbonate substrate, apolyimide substrate, a polyvinyl chloride substrate, a polyethylenesubstrate, a polypropylene substrate, a glycolated polyester (PETG)substrate, a polystyrene substrate, a silicon substrate, ceramicsubstrate, glass substrate or a polyester substrate (e.g., a 7millimeters thick polyester substrate). The electrically-insulatingsubstrate can have any suitable dimensions including, for example, awidth dimension of about 5 mm, a length dimension of about 27 mm and athickness dimension of about 0.5 mm.

First patterned conductor layer 504 can be formed of any suitableelectrically conductive material such as, for example, gold, palladium,carbon, silver, platinum, tin oxide, iridium, indium, or combinationsthereof (e.g., indium doped tin oxide). Moreover, any suitable techniqueor combination of techniques can be employed to form first patternedconductor layer 504 including, for example, sputtering, evaporation,electro-less plating, screen-printing, contact printing, laser ablationor gravure printing. A typical but non-limiting thickness for thepatterned conductor layer is in the range of 5 nanometers to 500nanometers.

Applicants note that conventional electrochemical-based analyte teststrips employ a working electrode along with an associatedcounter/reference electrode and enzymatic reagent layer to facilitate anelectrochemical reaction with an analyte of interest and, thereby,determine the presence and/or concentration of that analyte. Forexample, an electrochemical-based analyte test strip for thedetermination of glucose concentration in a fluid sample can employ anenzymatic reagent that includes the enzyme glucose oxidase and themediator ferricyanide (which is reduced to the mediator ferrocyanideduring the electrochemical reaction). Such conventional analyte teststrips and enzymatic reagent layers are described in, for example, U.S.Pat. Nos. 5,708,247; 5,951,836; 6,241,862; and 6,284,125; each of whichis hereby incorporated by reference herein to this application. In thisregard, the reagent layer employed in various embodiments providedherein can include any suitable sample-soluble enzymatic reagents, withthe selection of enzymatic reagents being dependent on the analyte to bedetermined and the bodily fluid sample. For example, if glucose is to bedetermined in a fluid sample, enzymatic reagent layer 506 can includeglucose oxidase or glucose dehydrogenase along with other componentsnecessary for functional operation.

In general, enzymatic reagent layer 506 includes at least an enzyme anda mediator. Examples of suitable mediators include, for example,ferricyanide, ferrocene, ferrocene derivatives, osmium bipyridylcomplexes, and quinone derivatives. Examples of suitable enzymes includeglucose oxidase, glucose dehydrogenase (GDH) using a pyrroloquinolinequinone (PQQ) co-factor, GDH using a nicotinamide adenine dinucleotide(NAD) co-factor, and GDH using a flavin adenine dinucleotide (FAD)co-factor. Enzymatic reagent layer 506 can be applied duringmanufacturing using any suitable technique including, for example,screen printing.

Applicants note that enzymatic reagent layer 506 may also containsuitable buffers (such as, for example, Tris HCl, Citraconate, Citrateand Phosphate), hydroxyethylcelulose [HEC], carboxymethylcellulose,ethycellulose and alginate, enzyme stabilizers and other additives asare known in the field.

Further details regarding the use of electrodes and enzymatic reagentlayers for the determination of the concentrations of analytes in abodily fluid sample, albeit in the absence of the phase-shiftmeasurement electrodes, bodily-fluid phase-shift sample chambers andsecond sample receiving channels analytical test strips and relatedmethods described herein, are in U.S. Pat. No. 6,733,655, which ishereby fully incorporated by reference herein to this application.

First and second patterned spacer layers 508 and 510 respectively can beformed of any suitable material including, for example, a 95 micrometersthick, double-sided pressure sensitive adhesive layer, a heat activatedadhesive layer, or a thermo-setting adhesive plastic layer. Firstpatterned spacer layer 508 can have, for example, a thickness in therange of from about 1 micron to about 500 microns, preferably betweenabout 10 microns and about 400 microns, and more preferably betweenabout 40 microns and about 600 microns.

Electrochemical-based analytical test strip 500 can be manufactured, forexample, by the sequential aligned formation of first patternedconductor layer 504, enzymatic reagent layer 506, first patterned spacerlayer 508, and second patterned spacer layer 510 ontoelectrically-insulating substrate 502. Any suitable techniques known toone skilled in the art can be used to accomplish such sequential alignedformation, including, for example, screen printing, photolithography,photogravure, chemical vapour deposition, sputtering, tape laminationtechniques and combinations thereof.

Analytical test strips according to embodiments can be configured, forexample, for operable electrical connection and use with the analyticaltest strip sample cell interface of a hand-held test meter as describedin co-pending patent application Ser. No. 13/250,525, which is herebyincorporated by reference herein to this application with a copyprovided in the Appendix.

It has been determined that a relationship exists between the reactanceof a fluid sample and the physical characteristic (e.g., hematocrit) ofthat sample. Electrical modeling of a bodily fluid sample (e.g., a wholeblood sample) as parallel capacitive and resistive components indicatesthat when an alternating signal such as, for example,alternating-current (AC) signal is forced through the bodily fluidsample, the phase shift of the alternating signal will be dependent onboth the frequency of the alternating signal voltage and the physicalcharacteristic (e.g., hematocrit) of the sample. Therefore, the physicalcharacteristic (e.g., hematocrit) of a bodily fluid sample can bemeasured by, for example, driving alternating signals of knownfrequencies through the bodily fluid sample and detecting their phaseshift. The phase-shift measurement electrodes of analytical test stripsof various embodiments described herein are particularly suitable foruse in such phase-shift measurements since the first and second phaseshift measurement electrodes are in direct contact with a bodily fluidsample present in the sample chamber. Moreover, a bodily fluid samplephysical characteristic (e.g., hematocrit) ascertained from a phaseshift measurement(s) can be employed to compensate for the effect ofphysical characteristic (e.g., hematocrit) during analyte determination.

Referring to FIGS. 3P-3T, electrochemical-based analytical test strip600 includes an electrically-insulating substrate 602, a first patternedconductor layer 604 disposed on the electrically-insulating substratelayer, an enzymatic reagent layer 606 (for clarity depicted in FIG. 3Ponly), a first patterned spacer layer 608, a second patterned conductorlayer 609, a second patterned spacer layer 610, and a top cover 611. Inthe embodiment of FIG. 3P, first pattered spacer layer 608 and secondpatterned spacer layer 610 are depicted as bi-layer structures. However,the first and second patterned spacer layers employed in variousembodiments provided herein can be unitary layers or any other suitablyformatted layer.

First patterned spacer layer 608 is configured such thatelectrochemical-based analytical test strip 600 also includes a firstsample-receiving channel 612, an analyte determination sample chamber614 and an analyte determination sample chamber vent 618 (not depictedin FIG. 3P but depicted with dashed lines in FIG. 3R). Analytedetermination sample chamber vent 618 is configured to aid in theintroduction of a bodily fluid sample into analyte determination samplechamber 614 via first sample-receiving channel 612.

Second patterned spacer layer 610 is configured to define a secondsample-receiving channel 620, a bodily fluid phase-shift sample chamber616 and a bodily fluid phase-shift chamber vent 622 (not depicted inFIG. 3P but depicted with dashed lines in FIG. 3S). Bodily fluidphase-shift chamber vent 622 is configured to aid in the introduction ofa bodily fluid sample into bodily fluid phase-shift sample chamber 616via second sample-receiving channel 620.

First patterned conductor layer 604 includes two working electrodes 628a and 628 b (depicted in FIGS. 3P and 3Q) and a reference electrode 630(also depicted in FIGS. 3P and 3Q). Second patterned conductor layer 609includes a first phase-shift measurement electrode 624 and a secondphase-shift measurement electrode 626 and is disposed above firstpatterned spacer layer 608 and embedded in the bi-layer structure ofsecond pattered spacer layer 610.

First sample-receiving channel 612 and analyte determination samplechamber 614 are isolated, both fluidically and electrically, from secondsample-receiving channel 620 and bodily fluid phase-shift sample chamber616 (see FIG. 3T in particular wherein the first and second patternedconductor layers are not depicted for clarity).

In the various embodiments of the test strip, there are two measurementsthat are made to a fluid sample deposited on the test strip. Onemeasurement is that of the concentration of the analyte (e.g. glucose)in the fluid sample while the other is that of physical characteristic(e.g., hematocrit) in the same sample. The measurement of the physicalcharacteristic (e.g., hematocrit) is used to modify or correct theglucose measurement so as to remove or reduce the effect of red bloodcells on the glucose measurements. Both measurements (glucose andhematocrit) can be performed in sequence, simultaneously or overlappingin duration. For example, the glucose measurement can be performed firstthen the physical characteristic (e.g., hematocrit); the physicalcharacteristic (e.g., hematocrit) measurement first then the glucosemeasurement; both measurements at the same time; or a duration of onemeasurement may overlap a duration of the other measurement. Eachmeasurement is discussed in detail as follow with respect to FIGS. 4A,4B and 5.

FIG. 4A is an exemplary chart of a test signal applied to test strip 100and its variations shown here in FIGS. 3A-3T. Before a fluid sample isapplied to test strip 100 (or its variants 400, 500, or 600), test meter200 is in a fluid detection mode in which a first test signal of about400 millivolts is applied between second working electrode and referenceelectrode. A second test signal of about 400 millivolts is preferablyapplied simultaneously between first working electrode (e.g., electrode12 of strip 100) and reference electrode (e.g., electrode 10 of strip100). Alternatively, the second test signal may also be appliedcontemporaneously such that a time interval of the application of thefirst test signal overlaps with a time interval in the application ofthe second test voltage. The test meter may be in a fluid detection modeduring fluid detection time interval T_(FD) prior to the detection ofphysiological fluid at starting time at zero. In the fluid detectionmode, test meter 200 determines when a fluid is applied to test strip100 (or its variants 400, 500, or 600) such that the fluid wets eitherthe first working electrode 12 or second working electrode 14 (or bothworking electrodes) with respect to reference electrode 10. Once testmeter 200 recognizes that the physiological fluid has been appliedbecause of, for example, a sufficient increase in the measured testcurrent at either or both of first working electrode 12 and secondworking electrode 14, test meter 200 assigns a zero second marker atzero time “0” and starts the test time interval T_(S). Test meter 200may sample the current transient output at a suitable sampling rate,such as, for example, every 1 milliseconds to every 100 milliseconds.Upon the completion of the test time interval T_(S), the test signal isremoved. For simplicity, FIG. 4A only shows the first test signalapplied to test strip 100 (or its variants 400, 500, or 600).

Hereafter, a description of how glucose concentration is determined fromthe known signal transients (e.g., the measured electrical signalresponse in nanoamperes as a function of time) that are measured whenthe test voltages of FIG. 4A are applied to the test strip 100 (or itsvariants 400, 500, or 600).

In FIG. 4A, the first and second test voltages applied to test strip 100(or its variants described herein) are generally from about +100millivolts to about +600 millivolts. In one embodiment in which theelectrodes include carbon ink and the mediator includes ferricyanide,the test signal is about +400 millivolts. Other mediator and electrodematerial combinations will require different test voltages, as is knownto those skilled in the art. The duration of the test voltages isgenerally from about 1 to about 5 seconds after a reaction period and istypically about 3 seconds after a reaction period. Typically, testsequence time T_(S) is measured relative to time t₀. As the voltage 401is maintained in FIG. 4A for the duration of T_(S), output signals aregenerated, shown here in FIG. 4B with the current transient 702 for thefirst working electrode 12 being generated starting at zero time andlikewise the current transient 704 for the second working electrode 14is also generated with respect to the zero time. It is noted that whilethe signal transients 702 and 704 have been placed on the samereferential zero point for purposes of explaining the process, inphysical term, there is a slight time differential between the twosignals due to fluid flow in the chamber towards each of the workingelectrodes 12 and 14 along axis L-L. However, the current transients aresampled and configured in the microcontroller to have the same starttime. In FIG. 4B, the current transients build up to a peak proximatepeak time Tp at which time, the current slowly drops off untilapproximately one of 2.5 seconds or 5 seconds after zero time. At thepoint 706, approximately at 5 seconds, the output signal for each of theworking electrodes 12 and 14 may be measured and added together.Alternatively, the signal from only one of the working electrodes 12 and14 can be doubled.

Referring back to FIG. 2B, the system drives a signal to measure orsample the output signals I_(E) from at least one the working electrodes(12 and 14) at any one of a plurality of time points or positions T₁,T₂, T₃, and T_(N). As can be seen in FIG. 4B, the time position can beany time point or interval in the test sequence T_(S). For example, thetime position at which the output signal is measured can be a singletime point T₁₅ at 1.5 seconds or an interval 708 (e.g., interval ˜10milliseconds or more depending on the sampling rate of the system)overlapping the time point T₂₈ proximate 2.8 seconds.

From knowledge of the parameters of the test strip (e.g., batchcalibration code offset and batch slope) for the particular test strip100 and its variations, the analyte (e.g., glucose) concentration can becalculated. Output transient 702 and 704 can be sampled to derivesignals I_(E) (by summation of each of the current I_(WE1) and I_(WE2)or doubling of one of I_(WE1) or I_(WE2)) at various time positionsduring the test sequence. From knowledge of the batch calibration codeoffset and batch slope for the particular test strip 100 and itsvariations in FIGS. 3B-3T, the analyte (e.g., glucose) concentration canbe calculated.

It is noted that “Intercept” and “Slope” are the values obtained bymeasuring calibration data from a batch of test strips. Typically,around 1500 strips are selected at random from the lot or batch.Physiological fluid (e.g., blood) from donors is spiked to variousanalyte levels, typically six different glucose concentrations.Typically, blood from 12 different donors is spiked to each of the sixlevels. Eight strips are given blood from identical donors and levels sothat a total of 12×6×8=576 tests are conducted for that lot. These arebenchmarked against actual analyte level (e.g., blood glucoseconcentration) by measuring these using a standard laboratory analyzersuch as Yellow Springs Instrument (YSI). A graph of measured glucoseconcentration is plotted against actual glucose concentration (ormeasured current versus YSI current) and a formula y=mx+c least squaresfitted to the graph to give a value for batch slope m and batchintercept c for the remaining strips from the lot or batch. Theapplicants have also provided methods and systems in which the batchslope is derived during the determination of an analyte concentration.The “batch slope”, or “Slope”, may therefore be defined as the measuredor derived gradient of the line of best fit for a graph of measuredglucose concentration plotted against actual glucose concentration (ormeasured current versus YSI current). The “batch intercept”, or“Intercept”, may therefore be defined as the point at which the line ofbest fit for a graph of measured glucose concentration plotted againstactual glucose concentration (or measured current versus YSI current)meets the y axis.

It is worthwhile here to note that the various components, systems andprocedures described earlier allow for applicants to provide an analytemeasurement system that heretofore was not available in the art. Inparticular, this system includes a test strip that has a substrate and aplurality of electrodes connected to respective electrode connectors.The system further includes an analyte meter 200 that has a housing, atest strip port connector configured to connect to the respectiveelectrode connectors of the test strip, and a microcontroller 300, shownhere in FIG. 2B. The microprocessor 300 is in electrical communicationwith the test strip port connector 220 to apply electrical signals orsense electrical signals from the plurality of electrodes.

Referring to FIG. 2B, details of a preferred implementation of meter 200where the same numerals in FIGS. 2A and 2B have a common description. InFIG. 2B, a strip port connector 220 is connected to the analogueinterface 306 by five lines including an impedance sensing line EIC toreceive signals from physical characteristic sensing electrode(s),alternating signal line AC driving signals to the physicalcharacteristic sensing electrode(s), reference line for a referenceelectrode, and signal sensing lines from respective working electrode 1and working electrode 2. A strip detection line 221 can also be providedfor the connector 220 to indicate insertion of a test strip. The analoginterface 306 provides four inputs to the processor 300: (1) realimpedance Z′; (2) imaginary impedance Z″; (3) signal sampled or measuredfrom working electrode 1 of the biosensor or I_(we1); (4) signal sampledor measured from working electrode 2 of the biosensor or I_(we2). Thereis one output from the processor 300 to the interface 306 to drive anoscillating signal AC of any value from 25 kHz to about 250 kHz orhigher to the physical characteristic sensing electrodes. A phasedifferential P (in degrees) can be determined from the real impedance Z′and imaginary impedance Z″ where:P=tan⁻¹ {Z″/Z′}  Eq. 3.1and magnitude M (in ohms and conventionally written as |Z|) from line Z′and Z″ of the interface 306 can be determined whereM=√{square root over ((Z′)²+(Z″)²)}  Eq. 3.2

In this system, the microprocessor is configured to: (a) apply a firstsignal to the plurality of electrodes so that a batch slope defined by aphysical characteristic of a fluid sample is derived and (b) apply asecond signal to the plurality of electrodes so that an analyteconcentration is determined based on the derived batch slope. For thissystem, the plurality of electrodes of the test strip or biosensorincludes at least two electrodes to measure the physical characteristicand at least two other electrodes to measure the analyte concentration.For example, the at least two electrodes and the at least two otherelectrodes are disposed in the same chamber provided on the substrate.Alternatively, the at least two electrodes and the at least two otherelectrodes are disposed in different chambers provided on the substrate.It is noted that for some embodiments, all of the electrodes aredisposed on the same plane defined by the substrate. In particular, insome of the embodiments described herein, a reagent is disposedproximate the at least two other electrodes and no reagent is disposedon the at least two electrodes. One feature of note in this system isthe ability to provide for an accurate analyte measurement within about10 seconds of deposition of a fluid sample (which may be a physiologicalsample) onto the biosensor as part of the test sequence.

As an example of an analyte calculation (e.g., glucose) for strip 100(FIG. 3A(1), 3A(2), or 3A(3) and its variants in FIGS. 3B-3T), it isassumed in FIG. 4B that the sampled signal value at 706 for the firstworking electrode 12 is about 1600 nanoamperes whereas the signal valueat 706 for the second working electrode 14 is about 1300 nanoamperes andthe calibration code of the test strip indicates that the Intercept isabout 500 nanoamperes and the Slope is about 18 nanoamperes/mg/dL.Glucose concentration G₀ can be thereafter be determined from Equation3.3 as follow:G ₀=[(I _(E))−Intercept]/Slope  Eq. 3.3where

I_(E) is a signal (proportional to analyte concentration) which is thetotal signal from all of the electrodes in the biosensor (e.g., forsensor 100, both electrodes 12 and 14 (or I_(we1)+I_(we2)));

I_(we1) is the signal measured for the first working electrode at theset sampling time;

I_(we2) is the signal measured for the second working electrode at theset sampling time;

Slope is the value obtained from calibration testing of a batch of teststrips of which this particular strip comes from;

Intercept is the value obtained from calibration testing of a batch oftest strips of which this particular strip comes from.

From Eq. 3.3; G₀=[(1600+1300)−500]/18 and therefore, G₀=133.33nanoamp˜133 mg/dL.

It is noted here that although the examples have been given in relationto a biosensor 100 which has two working electrodes (12 and 14 in FIG.3A(1)) such that the measured currents from respective workingelectrodes have been added together to provide for a total measuredcurrent I_(E), the signal resulting from only one of the two workingelectrodes can be multiplied by two in a variation of test strip 100where there is only one working electrode (either electrode 12 or 14).Instead of a total signal, an average of the signal from each workingelectrode can be used as the total measured current I_(E) for Equations3.3, 6, and 8-11 described herein, and of course, with appropriatemodification to the operational coefficients (as known to those skilledin the art) to account for a lower total measured current I_(E) than ascompared to an embodiment where the measured signals are added together.Alternatively, the average of the measured signals can be multiplied bytwo and used as I_(E) in Equations 3.3, 6, and 8-11 without thenecessity of deriving the operational coefficients as in the priorexample. It is noted that the analyte (e.g., glucose) concentration hereis not corrected for any physical characteristic (e.g., hematocritvalue) and that certain offsets may be provided to the signal valuesI_(we1) and I_(we2) to account for errors or delay time in theelectrical circuit of the meter 200. Temperature compensation can alsobe utilized to ensure that the results are calibrated to a referentialtemperature such as for example room temperature of about 20 degreesCelsius.

Now that a glucose concentration (G₀) can be determined from the signalI_(E), a description of applicant's technique to determine the physicalcharacteristic (e.g., hematocrit) of the fluid sample is provided inrelation to FIG. 5. In FIG. 5, the system 200 (FIG. 2) applies a firstoscillating input signal 800 at a first frequency (e.g., of about 25kilo-Hertz) to a pair of sensing electrodes. The system is also set upto measure or detect a first oscillating output signal 802 from thethird and fourth electrodes, which in particular involve measuring afirst time differential Δt₁ between the first input and outputoscillating signals. At the same time or during overlapping timedurations, the system may also apply a second oscillating input signal(not shown for brevity) at a second frequency (e.g., about 100kilo-Hertz to about 1 MegaHertz or higher, and preferably about 250 kiloHertz) to a pair of electrodes and then measure or detect a secondoscillating output signal from the third and fourth electrodes, whichmay involve measuring a second time differential Δt₂ (not shown) betweenthe first input and output oscillating signals. From these signals, thesystem estimates a physical characteristic (e.g., hematocrit) of thefluid sample based on the first and second time differentials Δt₁ andΔt₂. Thereafter, the system is able to derive a glucose concentration.The estimate of the physical characteristic (e.g., hematocrit) can bedone by applying an equation of the form

$\begin{matrix}{{HCT}_{EST} = \frac{\left( {{C_{1}\Delta\; t_{1}} - {C_{2}\Delta\; t_{2}} - C_{3}} \right)}{m_{1}}} & {{Eq}.\mspace{14mu} 4.1}\end{matrix}$where

each of C₁, C₂, and C₃ is an operational constant for the test strip and

m₁ represent a parameter from regressions data.

Details of this exemplary technique can be found in Provisional U.S.Patent Application Ser. No. 61/530,795 filed on Sep. 2, 2011, entitled,“Hematocrit Corrected Glucose Measurements for Electrochemical TestStrip Using Time Differential of the Signals”, which is herebyincorporated by reference.

Another technique to determine physical characteristic (e.g.,hematocrit) can be by two independent measurements of physicalcharacteristic (e.g., hematocrit). This can be obtained by determining:(a) the impedance of the fluid sample at a first frequency and (b) thephase angle of the fluid sample at a second frequency substantiallyhigher than the first frequency. In this technique, the fluid sample ismodeled as a circuit having unknown reactance and unknown resistance.With this model, an impedance (as signified by notation “|Z|”) formeasurement (a) can be determined from the applied voltage, the voltageacross a known resistor (e.g., the intrinsic strip resistance), and thevoltage across the unknown impedance Vz; and similarly, for measurement(b) the phase angle can be measured from a time difference between theinput and output signals by those skilled in the art. Details of thistechnique is shown and described in provisional patent application Ser.No. 61/530,808 filed Sep. 2, 2011, which is incorporated by reference.

Other suitable techniques for determining the physical characteristic(e.g., hematocrit, viscosity, temperature or density) of the fluidsample can also be utilized such as, for example, U.S. Pat. Nos.4,919,770, 7,972,861, US Patent Application Publication Nos.2010/0206749, 2009/0223834, or “Electric Cell—Substrate ImpedanceSensing (ECIS) as a Noninvasive Means to Monitor the Kinetics of CellSpreading to Artificial Surfaces” by Joachim Wegener, Charles R. Keese,and Ivar Giaever and published by Experimental Cell Research 259,158-166 (2000) doi:10.1006/excr.2000.4919, available online athttp://www.idealibrary.com; “Utilization of AC Impedance Measurementsfor Electrochemical Glucose Sensing Using Glucose Oxidase to ImproveDetection Selectivity” by Takuya Kohma, Hidefumi Hasegawa, DaisukeOyamatsu, and Susumu Kuwabata and published by Bull. Chem. Soc. Jpn.Vol. 80, No. 1, 158-165 (2007), all of these documents are incorporatedby reference.

Another technique to determine the physical characteristic (e.g.,hematorcrits, density, or temperature) can be obtained by knowing thephase difference (e.g., phase angle) and magnitude of the impedance ofthe sample. In one example, the following relationship is provided forthe estimate of the physical characteristic or impedance characteristicof the sample (“IC”):IC=M ² *y ₁ +M*y ₂ +y ₃ +P ² *y ₄ +P*y ₅   Eq. 4.2where

M represents a magnitude |Z| of a measured impedance in ohms);

P represents a phase difference between the input and output signals (indegrees)

y₁ is about −3.2e-08 and ±10%, 5% or 1% of the numerical value providedhereof (and depending on the frequency of the input signal, can bezero);

y₂ is about 4.1e-03 and ±10%, 5% or 1% of the numerical value providedhereof (and depending on the frequency of the input signal, can bezero);

y₃ is about −2.5e+01 and ±10%, 5% or 1% of the numerical value providedhereof

y₄ is about 1.5e-01 and ±10%, 5% or 1% of the numerical value providedhereof (and depending on the frequency of the input signal, can bezero); and

y₅ is about 5.0 and ±10%, 5% or 1% of the numerical value providedhereof (and depending on the frequency of the input signal, can bezero). The term “e” as used throughout this disclosure refers toscientific notation; e.g., 4.1e-03=4.1×10⁻³=0.0041.

It is noted here that where the frequency of the input AC signal is high(e.g., greater than 75 kHz) then the parametric terms y₁ and y₂ relatingto the magnitude of impedance M may be ±200% of the exemplary valuesgiven herein such that each of the parametric terms may include zero oreven a negative value. On the other hand, where the frequency of the ACsignal is low (e.g., less than 75 kHz), the parametric terms y₄ and y₅relating to the phase angle P may be ±200% of the exemplary values givenherein such that each of the parametric terms may include zero or even anegative value. It is noted here that a magnitude of H or HCT, as usedherein, is generally equal to the magnitude of IC. In one exemplaryimplementation, H or HCT is equal to IC as H or HCT is used herein thisapplication.

In another alternative implementation, Equation 4.3 is provided.Equation 4.3 is the exact derivation of the quadratic relationship,without using phase angles as in Equation 4.2.

$\begin{matrix}{{IC} = \frac{{- y_{2}} + {\sqrt{y_{2}^{2} - \left( {4{y_{3}\left( {y_{1} - M} \right)}} \right)}}}{2y_{1}}} & {{Eq}.\mspace{14mu} 4.3}\end{matrix}$where

IC is the Impedance Characteristic [%];

M is the magnitude of impedance [Ohm];

y₁ is about 1.2292e1 and ±10%, 5% or 1% of the numerical value providedhereof

y₂ is about −4.3431e2 and ±10%, 5% or 1% of the numerical value providedhereof

y₃ is about 3.5260e4 and ±10%, 5% or 1% of the numerical value providedhereof. As previously noted, the term “e” as used throughout thisdisclosure is intended to refer to scientific notation; e.g.,1.2292e1=1.2292×10¹=12.292.

By virtue of the various components, systems and insights providedherein, at least four techniques of determining an analyte concentrationfrom a fluid sample (which may be a physiological sample) (andvariations of such method) are achieved by applicants.

With reference to FIG. 6A(1), the method involves depositing a fluidsample (which may be a physiological sample) on a biosensor at step 904A(e.g., in the form of a test strip as show in FIG. 3A(1), 3A(2), or3A(3)-3T) that has been inserted into a meter (step 902A). Once themeter 200 is turned on, a voltage is applied to the strip 100 (or itsvariants 400, 500, or 600) and when the sample is deposited onto thetest chamber, the applied voltage physically transforms the analyte inthe sample into a different form due to the enzymatic reaction of theanalyte with the reagent in the test chamber. As the sample flows intothe capillary channel of the test cell, at least one physicalcharacteristic of the sample is obtained (step 908A). In particular, thestep of obtaining the physical characteristic (step 908A) may includeapplying a first signal to the sample to measure a physicalcharacteristic of the sample, while the step 906A of initiating anenzymatic reaction may involve driving a second signal to the sample,and the step of measuring (step 912A) may entail evaluating an outputsignal from the at least two electrodes at a point in time after thestart of the test sequence, in which a new batch slope is set (at step910A) as a function of at least the measured or estimated physicalcharacteristic (step 908A).

The setting of a new batch slope to derive a more accurate analyteconcentration measurement merits a discussion with reference to FIGS. 7and 6A(2). Applicants have found that the existing glucose test stripmade by LifeScan (marketed under the Ultra brand) has variations in thesignal output transients depending on the glucose concentration andhematocrit. These variations can be seen in FIG. 7 in which at highlevel of glucose (“High G”) or mid-level of glucose (“Mid-G”), thesignal transient varies distinctly as a function of the physicalcharacteristic (e.g., hematocrit) level and at low glucose level(“Lo-G”) the signal transient does not vary as distinctly as in theHigh-G or Mid-G as a function of hematocrit. Specifically, at the HighG, the signal transients 1000 a, 1002 a, and 1004 a (for 30%, 42% and55% Hct) maintain a generally consistent separation in signal outputover time after the peak at about 1.5 seconds after the start of thetest sequence. Similarly, at the Mid-G, the signal transients 1000 b,1002 b, and 1004 b (for 30%, 42%, and 55% Hct) maintains a consistentseparation in signal output over time after the peak at about 1.5seconds after the start of the test sequence. At the Low-G, the signaltransients 1000 c, 1002 c, and 1004 c (for 30%, 42%, and 55% Hct)generally converge together after the peak at about 1.5 seconds afterthe start of the test sequence.

Based on these observations, applicants have found that a relationshipexists between the batch slope of these test strips tested at the Lo-G,Mid-G, and Hi-G levels with respect to 30%, 42%, and 55% hematocritlevels. In particular, applicants have found that the batch slope forthese strips is generally curved with respect to hematocrit level, shownhere in FIG. 6A(2). In FIG. 6A(2), the batch slope declines in agenerally curved manner at low (e.g., 30%), medium (e.g., 42%), and high(e.g., 55%) hematocrits. As a consequence, by knowing the physicalcharacteristic of the sample (e.g., hematocrit) from Equation 4 above,the relationship in FIG. 6A(2) can be exploited to allow the slope inEquation 3.3 to accommodate the different levels of physicalcharacteristic (e.g., hematocrit) so as to achieve much more accurateglucose concentration measurements.

It should be noted that while slope “x” in FIG. 6A(2) appears to be alinear line, slope “x” is in fact a curved line and a curve has beenfitted for the relationship between physical characteristic (e.g.,hematocrit) and slope implicit in FIG. 6A(2). This fitted curve for FIG.6A(2) is found by applicants to be a second order equation of the form:NewSlope=aH ² bH+c  Eq. 5where

NewSlope is the derived or calculated new batch slope;

H is measured or estimated physical characteristic (e.g., hematocrit);

a is about 1.35e-6,

b is about −3.79e-4,

c is about 3.56e-2. As previously noted, the term “e” as used throughoutthis disclosure refers to scientific notation; e.g.,1.35e-6=1.35×10⁻⁶=0.00000135.

Equation 5 can be used instead of plotting against FIG. 6A(2), dependingon the available computing power of the processor. The viability of thisapproach can be seen here in FIG. 6A(3) which is a plot of a largenumber of test strips at different glucose ranges and hematocrit levelsversus percent bias by use of Equations 5 and 6. In FIG. 6A(3), it canbe seen that virtually all of the glucose concentration at differentglucose ranges (low, medium, and high) across about 30%, 42%, and about55% hematocrit have a bias of less than +10%.

Continuing on with the exemplary process of FIG. 6A(1), once thephysical characteristic is known at step 908A from Equation 4, such asfor example Hct ˜55%, the graph of FIG. 6A(2) is utilized to determinethe appropriate batch slope, designated here as “NewSlope” which isabout 0.019. Alternatively, Equation 5 can be utilized to derive theNewSlope from the measured or estimated physical characteristic (e.g.,hematocrit). This NewSlope (from FIG. 6A2 or Eq. 5) is used along withthe batch intercept (Intercept) of the particular batch of test strip inEquation 3.3, as set forth below in Equation 6.

$\begin{matrix}{G_{0} = {\left\lbrack \frac{I_{E} - {Intercept}}{x} \right\rbrack.}} & {{Eq}.\mspace{14mu} 6}\end{matrix}$where

G₀ represents an analyte concentration

I_(E) is a signal (proportional to analyte concentration) as measured atthe specified SamplingTime point;

x or “NewSlope” is the value obtained from the relationship in FIG. 6A2or from Eq. 5;

Intercept is the value obtained from calibration testing of a batch oftest strips of which this particular strip comes from.

Once the signal output I_(E) of the test chamber is measured at thedesignated time, typically at about 2.5 seconds or about 5 seconds, thesignal I_(E) is thereafter used with the NewSlope (of the batch) andIntercept (of the batch) in the calculation of the analyte concentration(in this case glucose) with Eq. 6 above. It is noted here that I_(E)could be a current from only one working electrode where the biosensorhas only one working electrode, a sum of current outputs from twoworking electrodes, a current from one working electrode multiplied bytwo or an average current from the two working electrodes. It should benoted that the step of applying the first signal and the driving of thesecond signal is in sequential order in that the order may be the firstsignal then the second signal or both signals overlapping in sequence;alternatively, the second signal first then the first signal or bothsignals overlapping in sequence. Alternatively, the applying of thefirst signal and the driving of the second signal may take placesimultaneously.

In the method, the step of applying of the first signal involvesdirecting an alternating signal provided by an appropriate power source(e.g., the meter 200) to the sample so that a physical characteristic ofthe sample is determined from an output of the alternating signal. Thephysical characteristic being detected may be one or more of viscosity,hematocrit, temperature or density. The directing step may includedriving first and second alternating signals at different respectivefrequencies in which a first frequency is lower than the secondfrequency. Preferably, the first frequency is at least one order ofmagnitude lower than the second frequency. As an example, the firstfrequency may be any frequency in the range of about 10 kHz to about 100kHz and the second frequency may be from about 250 kHz to about 1 MHz ormore. As used herein, the phrase “alternating signal” can have someportions of the signal alternating in polarity or all alternatingcurrent signal or an alternating current with a direct current offset oreven a multi-directional signal combined with a direct-current signal.

Although the method may specify only one sampling time point, the methodmay include sampling as many time points as required, such as, forexample, sampling the signal output continuously (e.g., at specifiedsampling time such as, every 10 milliseconds to 100 milliseconds) fromthe start of the test sequence until at least about 10 seconds after thestart and the results stored for processing near the end of the testsequence. Applicants note that the appropriate sampling time is measuredfrom the start of the test sequence but any appropriate datum may beutilized in order to determine when to sample the output signal. As apractical matter, the system can be programmed to sample the outputsignal at an appropriate time sampling interval during the entire testsequence such as for example, one sampling every 100 milliseconds oreven as little as about 1 milliseconds. In this variation, the sampledsignal output at the specified sampling time is the value used tocalculate the analyte concentration.

It is noted that in the preferred embodiments, the measure of a signaloutput for the glucose concentration is performed prior to themeasurement or estimation of the hematocrit. Alternatively, thehematocrit level can be measured or estimated prior to the measurementof the glucose concentration.

Thus, as another benefit of the teaching provided herein, a method ofdemonstrating increased accuracy of a test strip than heretofore isachieved. This method involves providing a batch of test strips,typically in a batch of at least about 1500 test strips (and in somecases up to 1 million test strips per batch), introducing a referentialsample containing a referential concentration of an analyte to each ofthe batch of test strips to initiate a test sequence (a “referentialsample” contains a “referential” or “known” concentration of an analyte,e.g. glucose). The method involves, reacting the analyte with a reagenton the test strip to cause a physical transformation of the analyteproximate the two electrodes, determining a physical characteristic ofthe referential sample, deriving a biosensor parameter (e.g., a batchslope) for the batch of test strips based on the determined physicalcharacteristics of the referential sample; sampling an electrical outputof the referential sample at a predetermined time point during the testsequence, and calculating an analyte concentration based on the derivedbiosensor parameter and sampled electrical output to provide for a finalanalyte concentration value for each of the batch of test strips suchthat at least 95% of the analyte concentration values of the batch oftest strips are within about ±15% of the referential analyteconcentration for the range of hematocrit from about 30% to about 55%,shown here in FIG. 6A(3) (an example of a “physical transformation”,specifically an “enzymatic reaction”, of an analyte to form a differentmaterial is the oxidation of glucose to D-glucono-δ-lactone and hydrogenperoxide by glucose oxidase (e.g. where the analyte is glucose and thereagent comprises glucose oxidase)).

In FIG. 6A3, experiments were performed to quantify the improvement inthe glucose measurements from the method noted above. The quantificationof the improvement can be shown by the “bias” at different levels ofhematocrit. The bias, which is an estimate of the relative error in theglucose measurement, was calculated for each glucose concentrationsdetermined with the method described in this example. The bias for eachglucose concentration was determined with equations of the form:Bias_(abs) =G _(calculated) −G _(reference) for G _(reference) less than100 mg/dL glucose and

${Bias}_{\%} = \frac{G_{calculated} - G_{reference}}{G_{reference}}$for G_(reference) greater than or equal to 100 mg/dL glucosewhere

Bias_(abs) is absolute bias,

Bias % is percent bias,

G_(calculated) is the glucose concentration determined by the methodherein and

G_(reference) is the reference glucose concentration.

As can be seen in FIG. 6A(3), most or virtually all of the glucoseconcentration using this technique are within about ±15% bias forhematocrit from about 30% hematocrit to about 55% hematocrit.Specifically, at region 1010A, only one glucose concentration greaterthan 100 mg/dL is outside the bias of −15%; at region 1012A ofintermediate hematocrit, a few glucose concentrations are outside thebias range of −15% and at region 1014A of high hematocrit, more glucoseconcentrations are dispersed outside the bias of −15% as compared toregions 1010A and 1012A.

With reference to FIG. 6B(1), another technique involves depositing afluid sample (which may be a physiological sample) on a biosensor atstep 904B (e.g., in the form of a test strip as show in FIG. 3A(1),3A(2), or 3A(3)-3T) that has been inserted into a meter (step 902B).Once the meter 200 is turned on, a voltage is applied to the strip 100(or its variants 400, 500, or 600) and when the sample is deposited ontothe test chamber, the applied voltage physically transforms the analytein the sample into a different form due to the enzymatic reaction of theanalyte with the reagent in the test chamber. As the sample flows intothe capillary channel of the test cell, at least one physicalcharacteristic of the sample is obtained (step 908B). In particular, thestep of obtaining or measuring the physical characteristic (step 908B)may include applying a first signal to the sample to derive a physicalcharacteristic of the sample, while the step 906B of initiating anenzymatic reaction (e.g., by applying electrical signals to the sampleand reagent) may involve driving a second signal to the sample, and thestep of measuring (step 912B) may entail measuring an output signal fromthe at least two electrodes at a point in time after the start of thetest sequence, in which a sampling time point is specified (at step910B) as a function of at least the measured or estimated physicalcharacteristic (step 908B).

In a variation of the method, the step of applying of the first signalinvolves directing an alternating signal provided by an appropriatepower source (e.g., the meter 200) to the sample so that a physicalcharacteristic of the sample is determined from an output of thealternating signal. The physical characteristic being detected may beone or more of viscosity, hematocrit or density of the sample (which maynot be physiological fluid such as, for example, calibration fluid). Thedirecting step may include driving first and second alternating signalsat different respective frequencies in which a first frequency is lowerthan the second frequency. Preferably, the first frequency is at leastone order of magnitude lower than the second frequency. As an example,the first frequency may be any frequency in the range of about 10 kHz toabout 100 kHz and the second frequency may be from about 250 kHz toabout 1 MHz or more. As used herein, the phrase “alternating signal” canhave some portions of the signal alternating in polarity or allalternating current signal or an alternating current with a directcurrent offset or even a multi-directional signal combined with adirect-current signal.

Once the physical characteristic of the sample is determined or obtainedfrom a suitable technique, the physical characteristic can be used tospecify a sampling time point at which point during the test sequencethe output signal of the test chamber could be measured. In particular,applicants have found a relationship between the physical characteristicand the sampling time point, as shown here in FIG. 7. This relationshiphas been further explored such that applicants were able to derive adirect relationship between the sampling time point of the sample andthe physical characteristic of the sample (e.g., hematocrit), shown herein FIG. 6B(2). As a consequence, by knowing the physical characteristicof the sample (e.g., hematocrit) from Equation 4 above, the relationshipin FIG. 6B(2) can be exploited to allow the sampling time point to bespecified to accommodate the different levels of physical characteristic(e.g., hematocrit) so as to achieve much more accurate glucoseconcentration measurements than heretofore.

Referring back to FIG. 7, it can be seen that as the analyteconcentration (proportional to the signal output) increases, the peak ofthe high glucose concentration (denoted by 1002 a, 1004 a, and 1006 a)is shifted to the right as compared to the medium glucose concentration(denoted by 1002 b, 1004 b, and 1006 b). Similarly, the peak of themedium glucose concentration is further to the right of FIG. 7 ascompared to low glucose concentration (denoted by 1002 c, 1004 c, and1006 c). It can also be seen here that the steady-state of the lowglucose concentrations (1002 c, 1004 c, and 1006 c) is reached earlierthan the medium glucose concentrations (1002 b, 1004 b, and 1006 b).This pattern is repeated for high glucose concentration (1002 a, 1004 a,and 1006 b) as compared to medium glucose concentrations.

From data in FIG. 7, applicants were able to derive a second degreerelationship between the sensed physical characteristic and the samplingtime, shown here in FIG. 6B(2). In FIG. 6B(2), a curve is fitted tohematocrit values at about 30%, 42% and about 55% and glucose values forthese ranges of hematocrits (from FIG. 7). This fitted curve is found byapplicants to be an equation of the form:SamplingTime=x ₁ H ^(x) ² +x ₃  Eq. 7where

“Sampling Time” is designated (for convenience) as a time point from thestart of the test sequence at which to sample the output signal of thetest strip,

H represents the physical characteristic of the sample;

x₁ is about 4.3e5;

x₂ is about −3.9; and

x₃ is about 4.8. As previously noted, the term “e” as used throughoutthis disclosure refers to scientific notation; e.g.,4.3e5=4.3×10⁵=430,000.

Although the method may specify only one sampling time point, the methodmay include sampling as many time points as required, such as, forexample, sampling the signal output continuously (e.g., at specifiedsampling time such as, every 10 milliseconds to 100 milliseconds) fromthe start of the test sequence until at least about 10 seconds after thestart and the results stored for processing near the end of the testsequence. Applicants note that the appropriate sampling time is measuredfrom the start of the test sequence but any appropriate datum may beutilized in order to determine when to sample the output signal. As apractical matter, the system can be programmed to sample the outputsignal at an appropriate time sampling interval during the entire testsequence such as for example, one sampling every 100 milliseconds oreven as little as about 1 milliseconds. In this variation, the sampledsignal output at the specified sampling time is the value used tocalculate the analyte concentration.

Referring back to FIG. 6B(1), the method can now determine the analyteconcentration (step 914B) based on the measured signal (step 912B)sampled at the specified time point, which specified time point is afunction of the obtained or measured or estimated physicalcharacteristic of the sample (e.g., from plot of FIG. 6B(2) or Equation7). That is, once the sampling time T has been specified from Equation7, the method provides for measuring the output signal from the testchamber (e.g., chamber 92) so that the sampled output can be utilizedwith Equation 3.3 to provide for an analyte (e.g., glucose)concentration.

Alternatively, a look-up table, represented exemplarily here withreference to Table 1 can also be utilized in place of Equation 7 or inaddition to Equation 7 to specify an appropriate sampling time point. InTable 1, the value of the physical characteristic is used by theprocessor of the system to look up the appropriate time at which thesignal output of the biosensor is sampled or measured to determine theanalyte concentration. For example, once the physical characteristic hasbeen determined, in this case about 33% hematocrit, the time at whichthe signal output of the biosensor 100 is utilized in determining theanalyte concentration can be gleaned from Table 1, which shows that thetime at which the system must sample the signal output is atapproximately 5.32 seconds after the start of the test sequence.

TABLE 1 Physical Characteristic Sampling Time T (e.g., Hematocrit %)(seconds) 30 5.56 31 5.46 32 5.38 33 5.32 34 5.26 35 5.2 36 5.16 37 5.1238 5.08 39 5.06 40 5.02 41 5 42 5 43 4.98 44 4.96 45 4.96 46 4.94 474.92 48 4.92 49 4.9 50 4.9 51 4.9 52 4.88 53 4.88 54 4.88 55 4.86

It should be noted that the step of applying the first signal and thedriving of the second signal is in sequential order in that the ordermay be the first signal then the second signal or both signalsoverlapping in sequence; alternatively, the second signal first then thefirst signal or both signals overlapping in sequence. Alternatively, theapplying of the first signal and the driving of the second signal maytake place simultaneously.

It is noted that in the preferred embodiments, the measurement of asignal output for the glucose concentration is performed prior to theestimation of the physical characteristic (e.g., hematocrit).Alternatively, the physical characteristic (e.g., hematocrit) level canbe estimated, measured, or obtained prior to the measurement of theglucose concentration.

Thus, as another benefit of the teaching provided herein, a method ofdemonstrating increased accuracy of a test strip than heretofore isachieved. This method involves providing a batch of test strips,typically in a batch of at least about 1500 test strips (and in somecases, up to 1 million test strips per batch), introducing a referentialsample containing a referential concentration of an analyte to each ofthe batch of test strips to initiate a test sequence. The methodinvolves reacting the analyte to cause a physical transformation of theanalyte with the reagent between the two electrodes, determining aphysical characteristic of the referential sample, estimating theanalyte concentration, sampling an electrical output of the referentialsample at a specified time point during the test sequence defined by themeasured or estimated physical characteristic of the sample and theestimated analyte concentration, and calculating an analyteconcentration based on the specified sampling time such that at least95% of the analyte concentration values of the batch of test strips arewithin about 25% of the referential analyte concentration for the rangeof hematocrit from about 30% to about 55%, shown here in FIG. 6B(3).

As can be seen in FIG. 6B(3), most or virtually all of the glucoseconcentration using this technique are within about ±25% bias forhematocrit from about 30% hematocrit to about 55% hematocrit.Specifically, at region 1010B, only one glucose concentration greaterthan 100 mg/dL is outside the bias of −15%; at region 1012B ofintermediate hematocrit, a few glucose concentrations are outside thebias range of about −25% and at region 1014B of high hematocrit, moreglucose concentrations are dispersed outside the bias of about −25% ascompared to regions 1010B and 1012B.

Yet another technique can be understood with reference to FIG. 6C(1).This technique involves depositing a fluid sample (which may be aphysiological sample) on a biosensor at step 904C (e.g., in the form ofa test strip as show in FIG. 3A(1), 3A(2), or 3A(3)-3T) that has beeninserted into a meter (step 902C). Once the meter 200 is turned on, avoltage is applied to the strip 100 (or its variants 400, 500, or 600)and when the sample is deposited onto the test chamber, the appliedvoltage physically transforms the analyte in the sample into a differentform due to the enzymatic reaction of the analyte with the reagent inthe test chamber. As the sample flows into the capillary channel of thetest cell, at least one physical characteristic of the sample isobtained (step 908C). In particular, the step of obtaining or measuringthe physical characteristic (step 908C) may include applying a firstsignal to the sample to derive a physical characteristic of the sample,while the step 906C of initiating an enzymatic reaction (e.g., byapplying signals to the sample and reagent) may involve driving a secondsignal to the sample, and the step of measuring (step 912C) may entailmeasuring an output signal from the at least two electrodes at a pointin time after the start of the test sequence, in which a sampling timepoint is specified (at step 909) and a batch slope (step 910C) isderived and as a function of at least the measured or estimated physicalcharacteristic (step 908C).

In a variation of the method, the step of applying of the first signalinvolves directing an alternating signal provided by an appropriatepower source (e.g., the meter 200) to the sample so that a physicalcharacteristic of the sample is determined from an output of thealternating signal. The physical characteristic being detected may beone or more of viscosity, hematocrit or density. The directing step mayinclude driving first and second alternating signal at differentrespective frequencies in which a first frequency is lower than thesecond frequency. Preferably, the first frequency is at least one orderof magnitude lower than the second frequency. As an example, the firstfrequency may be any frequency in the range of about 10 kHz to about 100kHz and the second frequency may be from about 250 kHz to about 1 MHz ormore. As used herein, the phrase “alternating signal” can have someportions of the signal alternating in polarity or all alternatingcurrent signal or an alternating current with a direct current offset oreven a multi-directional signal combined with a direct-current signal.

Once the physical characteristic of the sample is determined or obtainedfrom a suitable technique, the physical characteristic can be used tospecify a sampling time point (step 909) at which point during the testsequence the output signal of the test chamber could be measured. Inparticular, applicants have found a relationship between the physicalcharacteristic and the sampling time point, as shown here in FIG. 7.This relationship has been further explored such that applicants wereable to derive a direct relationship between the sampling time point ofthe sample and the physical characteristic of the sample (e.g.,hematocrit), shown here in FIG. 6C(2). As a consequence, by knowing thephysical characteristic of the sample (e.g., hematocrit) from Equation 4above, the relationship in FIG. 6C(2) can be exploited to allow thesampling time point to be specified to accommodate the different levelsof physical characteristic (e.g., hematocrit) so as to achieve much moreaccurate analyte (e.g., glucose) concentration measurements.

Referring back to FIG. 7, it can be seen that as the analyteconcentration (proportional to the signal output) increases, the peak ofthe high glucose concentration (denoted by 1002 a, 1004 a, and 1006 a)is shifted to the right as compared to the medium glucose concentration(denoted by 1002 b, 1004 b, and 1006 b). Similarly, the peak of themedium glucose concentration is further to the right of FIG. 7 ascompared to low glucose concentration (denoted by 1002 c, 1004 c, and1006 c). It can also be seen here that the steady-state of the lowglucose concentrations (1002 c, 1004 c, and 1006 c) is reached earlierthan the medium glucose concentrations (1002 b, 1004 b, and 1006 b).This pattern is repeated for high glucose concentration (1002 a, 1004 a,and 1006 b) as compared to medium glucose concentrations.

Although the method may specify only one sampling time point, the methodmay include sampling as many time points as required, such as, forexample, sampling the signal output continuously (e.g., at specifiedsampling time such as, every one millisecond to 100 milliseconds) fromthe start of the test sequence until at least about 10 seconds after thestart and the results stored for processing near the end of the testsequence. Applicants note that the appropriate sampling time is measuredfrom the start of the test sequence but any appropriate datum may beutilized in order to determine when to sample the output signal. As apractical matter, the system can be programmed to sample the outputsignal at an appropriate time sampling interval during the entire testsequence such as for example, one sampling every 100 milliseconds oreven as little as about 1 milliseconds. In this variation, the sampledsignal output at the specified sampling time T is the value used tocalculate the analyte concentration.

Alternatively, a look-up table, represented exemplarily here withreference to Table 1 can also be utilized in place of Equation 7 or inaddition to Equation 7 to determine the appropriate sampling time pointT. In Table 1, the value of the physical characteristic is used by theprocessor of the system to look up the appropriate time at which thesignal output of the biosensor is sampled or measured to determine theanalyte concentration. For example, once the physical characteristic hasbeen determined, in this case about 33% hematocrit, the time at whichthe signal output of the biosensor 100 is utilized in determining theanalyte concentration can be gleaned from Table 1, which shows that thetime at which the system must sample the signal output is atapproximately 5.32 seconds after the start of the test sequence.

To further improve the accuracy of the results, the method may entailevaluating an output signal from the at least two electrodes at a pointin time after the start of the test sequence, in which a new batch slopeis derived (at step 910C) as a function of at least the measured orestimated physical characteristic (step 908C).

The setting of a new batch slope (step 910C) to derive a more accurateanalyte concentration measurement merits a discussion with reference toFIGS. 7 and 6C(3). Applicants have further found that anotherrelationship exists between the batch slope of these test strips testedat the Lo-G, Mid-G, and Hi-G levels with respect to 30%, 42%, and 55%hematocrit levels of FIG. 7. In particular, applicants have found thatthe batch slope for these strips is generally curved with respect tohematocrit level, shown here in FIG. 6C(3). In FIG. 6C(3), the batchslope declines in a generally curved manner at low (e.g., 30%), medium(e.g., 42%), and high (e.g., 55%) hematocrits. As a consequence, byknowing the physical characteristic of the sample (e.g., hematocrit)from Equation 4 above, the relationship in FIG. 6C(3) can be exploitedto allow the slope in Equation 3.3 to be calculated for the differentlevels of physical characteristic (e.g., hematocrit) so as to achievemuch more accurate glucose concentration measurements.

It should be noted that while slope “x” in FIG. 6C(3) appears to be alinear line, slope “x” is in fact a curved line and a curve has beenfitted for the relationship between physical characteristic (e.g.,hematocrit) and slope implicit in FIG. 6C(3). This fitted curve for FIG.6C(3) was found by applicants and defined in Equation 5 previously. Itis noted that due to an adjustment at which the sampling time was takenfor the embodiment described herein, Equation 5 is preferably used withthe following coefficients: a is about −1.98e-6; b is about −2.87e-5;and c is about 2.67e-2 and ±10% for each of the magnitudes provided. Aspreviously noted, the term “e” as used throughout this disclosure refersto scientific notation; e.g., −1.98e-06=−1.98×10⁻⁶=−0.00000198. Theadjustment stems from the fact that the new coefficients (a, b, and c)are calculated at different sampling time, whereas before, the samplingtime was fixed at about 5 seconds. This will cause coefficients a, b andc to be different in order to maximize accuracy. If one skilled in theart were to use the same coefficients a, b and c as before, such skilledperson can still obtain the analyte concentration but the resultingestimates will deteriorate. The driver here is that slope of glucoserelation to haematocrit is changing within the signal transient, sodifferent slopes are needed as the sampling time is varied between about3.5 seconds to about 6 seconds.

It is noted that Equation 5 can be used instead of plotting against FIG.6C(3), depending on the available computing power of the processor.Continuing on with the exemplary process of FIG. 6C(1), once thephysical characteristic is known at step 908C from Equation 4, such asfor example Hct ˜55%, the graph of FIG. 6C(3) or Equation 5 is utilizedto determine the appropriate batch slope, designated here as “NewSlope”which is about 0.019. Alternatively, Equation 5 can be utilized toderive the NewSlope from the measured or estimated physicalcharacteristic (e.g., hematocrit). This NewSlope (from FIG. 6C3 or Eq.5) is used along with the batch intercept (Intercept) with the samplingtime relationship that applicants have found, as set forth previouslywith respect to Eq. 5.

Referring back to FIG. 6C(1), the method can now determine the analyteconcentration (step 914C) based on a new batch slope (i.e., NewSlope)derived from the measured or estimated physical characteristic of thesample (at step 910C) along with the measured signal (step 912C) sampledat the specified time point (at step 909), which specified time point isa function of the obtained or measured or estimated physicalcharacteristic of the sample (e.g., from plot of FIG. 6C(2) or Equation5). That is, once the sampling time has been specified from Equation 5and the NewSlope from Equation 6, the method provides for measuring theoutput signal from the test chamber (e.g., chamber 92) so that thesampled output can be utilized with Equation 3.3 to provide for ananalyte (e.g., glucose) concentration.

In one embodiment, the analyte (e.g., glucose) concentration isdetermined based on a modified form of Equation 3.3, delineated here asEquation 8:

$\begin{matrix}{G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{NewSlope} \right\rbrack} & {{Eq}.\mspace{14mu} 8}\end{matrix}$where

G₀ represents an analyte concentration;

I_(E) is a signal (proportional to analyte concentration) as measured atthe specified SamplingTime point;

“NewSlope” is the value obtained from the relationship in FIG. 6C(3) orfrom Eq. 5;

Intercept is the value obtained from calibration testing of a batch oftest strips of which this particular strip comes from.

Once the signal output I_(E) of the test chamber is measured at thespecified time (step 909) at any point between about 3 seconds to about8 seconds, the signal I_(E) is thereafter used with the NewSlope (of thebatch) and Intercept (of the batch) in the calculation of the analyteconcentration (in this case glucose) with Eq. 8 above. Interceptcomprises a manufacturing parameter for a batch of biosensors, and inthe embodiments described herein, Intercept typically varies from about0.7 to about 0.6. It should be noted that the step of applying the firstsignal and the driving of the second signal is in sequential order inthat the order may be the first signal then the second signal or bothsignals overlapping in sequence; alternatively, the second signal firstthen the first signal or both signals overlapping in sequence.Alternatively, the applying of the first signal and the driving of thesecond signal may take place simultaneously.

It is noted that in the preferred embodiments, the measurement of asignal output for the analyte (e.g., glucose) concentration is performedprior to the estimation of the physical characteristic (e.g.,hematocrit). Alternatively, the physical characteristic (e.g.,hematocrit) level can be estimated, measured, or obtained prior to themeasurement of the analyte (e.g., glucose) concentration.

Thus, as another benefit of the teaching provided herein, a method ofdemonstrating increased accuracy of a test strip is heretofore isachieved. This method involves providing for a batch of test strips,typically in a batch of at least about 1500 test strips (and in somecases up to 1 million test strips per batch), introducing a referentialsample containing a referential concentration of an analyte to each ofthe batch of test strips to initiate a test sequence. The methodinvolves reacting the analyte to cause a physical transformation of theanalyte with the reagent between the two electrodes, determining aphysical characteristic of the referential sample, deriving a batchslope for the test strip, sampling an electrical output of thereferential sample at a specified time point during the test sequencedefined by the measured or estimated physical characteristic of thesample, and calculating an analyte concentration based on the dictatedtime and the derive batch slope such that at least 95% of the analyteconcentration values of the batch of test strips are within about ±15%of the referential analyte concentration for the range of hematocritfrom about 30% to about 55%, shown here in FIG. 6C(4).

As can be seen in FIG. 6C(4), most or virtually all of the glucoseconcentration using this technique are within about ±15% bias forhematocrit from about 30% hematocrit to about 55% hematocrit.Specifically, at region 1010C, none of the glucose concentration greaterthan 100 mg/dL is outside the bias of −15%; at region 1012C ofintermediate hematocrit, a few glucose concentrations are outside thebias range of 15% and at region 1014C of high hematocrit, a few moreglucose concentrations are dispersed outside the bias of 15% as comparedto regions 1010C and 1012C.

Yet a further technique can be understood with reference to FIG. 6D(1).This technique involves depositing a fluid sample (which may be aphysiological sample) on a biosensor at step 904 (e.g., in the form of atest strip as show in FIG. 3A(1), 3A(2), or 3A(3)-3T) that has beeninserted into a meter (step 902). Once the meter 200 is turned on, avoltage is applied to the strip 100 (or its variants 400, 500, or 600)and when the sample is deposited onto the test chamber, the appliedsignal physically transforms the analyte in the sample into a differentform due to the enzymatic reaction of the analyte with the reagent inthe test chamber. As the sample flows into the capillary channel of thetest cell, at least one physical characteristic of the sample isobtained (step 908) along with estimate of the analyte concentration(step 910). From the obtained physical characteristic (step 908) andestimated analyte concentration (step 910), a sampling time point isdefined at which the signal output from the sample during the testsequence is measured (at step 914) and used for calculating the analyteconcentration in step 916. In particular, the step of obtaining thephysical characteristic (step 908) may include applying a first signalto the sample to measure a physical characteristic of the sample, whilethe step 906 of initiating an enzymatic reaction may involve driving asecond signal to the sample, and the step of measuring (step 914) mayentail evaluating an output signal from the at least two electrodes at apoint in time after the start of the test sequence, in which the pointin time is set (at step 912) as a function of at least the measured orestimated physical characteristic (step 908) and estimated analyteconcentration (step 910).

The determination of the appropriate point in time as a function of themeasured or estimated physical characteristic(s) in step 912 can bedetermined by the use of a look-up table programmed into themicroprocessor of the system. For example, a look-up table may beprovided that allows for the system to select the appropriate samplingtime for the analyte (e.g., glucose or ketone) with measured or knownphysical characteristic (e.g., hematocrit or viscosity) of the sample.

In particular, an appropriate sampling time point may be based on anearly estimation of the analyte and the measured or known physicalcharacteristic to arrive at the appropriate sampling time that gives thelowest error or bias as compared to referential values. In thistechnique, a look up table is provided in which the defined samplingtime point is correlated to (a) the estimated analyte concentration and(b) the physical characteristic of the sample. For example, Table 2 maybe programmed into the meter to provide a matrix in which qualitativecategories (low, medium, and high glucose) of the estimated analyte formthe main column and the qualitative categories (low, medium, and high)of the measured or estimated physical characteristic form the headerrow. In the second column, t/Hct is a value determined experimentally ofthe time shift per % hematocrit difference from nominal hematocrit of42%. As one example, for 55% hematocrit at “Mid-Glucose” would indicatea time shift of (42−55)*90=−1170 ms. The time of −1170 milliseconds isadded to the original test time of about 5000 milliseconds giving(5000−1170=3830 milliseconds) ˜3.9 seconds.

TABLE 2 Sampling Time Sampling Time Sampling Time Point T for Lo HctPoint T for Mid Hct Point T for High Hct Estimated t/Hct (from start oftest (from start of test (from start of test Analyte (in milliseconds)sequence, in seconds) sequence, in seconds) sequence, in seconds)Lo-Glucose 40 5.5 5 4.5 Mid-Glucose 90 6.1 5 3.9 Hi-Glucose 110 6.3 53.6

The time T at which the system should be sampling the output signal ofthe biosensor is based on both the qualitative category of the estimatedanalyte and measured or estimated physical characteristic and ispredetermined based on regression analysis of a large sample size ofactual physiological fluid samples. Applicants note that the appropriatesampling time is measured from the start of the test sequence but anyappropriate datum may be utilized in order to determine when to samplethe output signal. As a practical matter, the system can be programmedto sample the output signal at an appropriate time sampling intervalduring the entire test sequence such as for example, one sampling every100 milliseconds or even as little as about 1 milliseconds. By samplingthe entire signal output transient during the test sequence, the systemcan perform all of the needed calculations near the end of the testsequence rather than attempting to synchronize the sampling time withthe set time point, which may introduce timing errors due to systemdelay.

Applicants hereafter will discuss the look-up Table 2 in relation to theparticular analyte of glucose in physiological fluid samples.Qualitative categories of blood glucose are defined in the first columnof Table 2 in which low blood glucose concentrations of less than about70 mg/dL are designated as “Lo-Glucose”; blood glucose concentrations ofhigher than about 70 mg/dL but less than about 250 mg/dL are designatedas “Mid-Glucose”; and blood glucose concentrations of higher than about250 mg/dL are designated as “Hi-Glucose”.

During a test sequence, an “Estimated Analyte” can be obtained bysampling the signal at a convenient time point, typically at fiveseconds during a typical 10 seconds test sequence. The measurementsampled at this five second time point allows for an accurate estimateof the analyte (in this case blood glucose). The system may then referto a look-up table (e.g., Table 2) to determine when to measure thesignal output from the test chamber at a specified sampling time T basedon two criteria: (a) estimated analyte and (b) qualitative value of thephysical characteristic of the sample. For criteria (b), the qualitativevalue of the physical characteristic is broken down into threesub-categories of Low Hct, Mid Hct and High Hct. Thus, in the event thatthe measured or estimated physical characteristic (e.g., hematocrit) ishigh (e.g., greater than 46%) and the estimated glucose is also high,then according to Table 2, the test time for the system to measure thesignal output of test chamber would be about 3.6 seconds. On the otherhand, if the measured hematocrit is low (e.g., less than 38%) and theestimated glucose is low then according to Table 2, the test time T forthe system to measure the signal output of test chamber would be about5.5 seconds.

Once the signal output I_(T) of the test chamber is measured at thedesignated time (which is governed by the measured or estimated physicalcharacteristic), the signal I_(T) is thereafter used in the calculationof the analyte concentration (in this case glucose) with Equation 9below.

$\begin{matrix}{G_{0} = \left\lbrack \frac{I_{T} - {Intercept}}{Slope} \right\rbrack} & {{Eq}.\mspace{14mu} 9}\end{matrix}$where

G₀ represents an analyte concentration;

I_(T) represents a signal (proportional to analyte concentration)determined from the sum of the end signals measured at a specifiedsampling time T, which may be the total current measured at thespecified sampling time T;

Slope represents the value obtained from calibration testing of a batchof test strips of which this particular strip comes from and istypically about 0.02; and

Intercept represents the value obtained from calibration testing of abatch of test strips of which this particular strip comes from and istypically from about 0.6 to about 0.7.

It should be noted that the step of applying the first signal and thedriving of the second signal is sequential in that the order may be thefirst signal then the second signal or both signals overlapping insequence; alternatively, the second signal first then the first signalor both signals overlapping in sequence. Alternatively, the applying ofthe first signal and the driving of the second signal may take placesimultaneously.

In the method, the step of applying of the first signal involvesdirecting an alternating signal provided by an appropriate power source(e.g., the meter 200) to the sample so that a physical characteristic ofthe sample is determined from an output of the alternating signal. Thephysical characteristic being detected may be one or more of viscosity,hematocrit or density. The directing step may include driving first andsecond alternating signal at different respective frequencies in which afirst frequency is lower than the second frequency. Preferably, thefirst frequency is at least one order of magnitude lower than the secondfrequency. As an example, the first frequency may be any frequency inthe range of about 10 kHz to about 100 kHz and the second frequency maybe from about 250 kHz to about 1 MHz or more. As used herein, the phrase“alternating signal” or “oscillating signal” can have some portions ofthe signal alternating in polarity or all alternating current signal oran alternating current with a direct current offset or even amulti-directional signal combined with a direct-current signal.

Further refinements of Table 2 based on additional investigations of thetechnique allowed applicants to devise Table 3, shown below.

TABLE 3 Sampling Time S to Estimated G and Measured or EstimatedPhysical Characteristic Estimated Measured or Estimated Physical GCharacteristic (e.g., HCT [%]) [mg/dL] 24 27 30 33 36 39 42 45 48 51 5457 60 25 4.6 4.6 4.5 4.4 4.4 4.4 4.3 4.3 4.3 4.2 4.1 4.1 4.1 50 5 4.94.8 4.7 4.7 4.6 4.5 4.4 4.3 4.2 4.1 4 4 75 5.3 5.3 5.2 5 4.9 4.8 4.7 4.54.4 4.3 4.1 4 3.8 100 5.8 5.6 5.4 5.3 5.1 5 4.8 4.6 4.4 4.3 4.1 3.9 3.7125 6.1 5.9 5.7 5.5 5.3 5.1 4.9 4.7 4.5 4.3 4.1 3.8 3.6 150 6.4 6.2 5.95.7 5.5 5.3 5 4.8 4.6 4.3 4 3.8 3.5 175 6.6 6.4 6.2 5.9 5.6 5.4 5.2 4.94.6 4.3 4 3.7 3.4 200 6.8 6.6 6.4 6.1 5.8 5.5 5.2 4.9 4.6 4.3 4 3.7 3.4225 7.1 6.8 6.5 6.2 5.9 5.6 5.3 5 4.7 4.3 4 3.6 3.2 250 7.3 7 6.7 6.4 65.7 5.3 5 4.7 4.3 4 3.6 3.2 275 7.4 7.1 6.8 6.4 6.1 5.8 5.4 5 4.7 4.3 43.5 3.2 300 7.5 7.1 6.8 6.5 6.2 5.8 5.5 5.1 4.7 4.3 4 3.5 3.1 w325 7.67.3 6.9 6.5 6.2 5.8 5.5 5.1 4.7 4.3 3.9 3.5 3.1 350 7.6 7.3 7 6.6 6.25.8 5.5 5.1 4.7 4.3 3.9 3.5 3.1 375 7.7 7.3 7 6.6 6.2 5.8 5.5 5.1 4.74.3 3.9 3.5 3.1 400 7.7 7.3 6.9 6.5 6.2 5.8 5.4 5 4.7 4.3 3.9 3.5 3.1425 7.6 7.3 6.9 6.5 6.2 5.8 5.4 5 4.6 4.3 3.8 3.5 3.1 450 7.6 7.2 6.86.4 6.1 5.7 5.3 5 4.6 4.3 3.8 3.5 3.1 475 7.4 7.1 6.7 6.4 6 5.6 5.3 4.94.6 4.2 3.8 3.5 3.1 500 7.3 7 6.6 6.2 5.9 5.5 5.2 4.9 4.5 4.1 3.8 3.53.2 525 7.1 6.8 6.5 6.1 5.8 5.5 5.1 4.8 4.4 4.1 3.8 3.5 3.2 550 7 6.76.3 5.9 5.6 5.3 5 4.7 4.4 4.1 3.8 3.5 3.2 575 6.8 6.4 6.1 5.8 5.5 5.24.9 4.6 4.3 4.1 3.8 3.5 3.4 600 6.5 6.2 5.9 5.6 5.3 5 4.7 4.5 4.3 4 3.83.6 3.4

As in Table 2, a measured or estimated physical characteristic is usedin Table 3 along with an estimated analyte concentration to derive atime S at which the sample is to be measured. For example, if themeasured charactertistic is about 30% and the estimated glucose (e.g.,by sampling at about 2.5 to 3 seconds) is about 350, the time at whichthe microcontroller should sample the fluid is about 7 seconds. Inanother example, where the estimated glucose is about 300 mg/dL and themeasured or estimated physical characteristic is 60%, specified samplingtime would be about 3.1 seconds.

For the embodiments utilized with Table 3, the estimated glucoseconcentration is provided with an equation:

$\begin{matrix}{G_{est} = \frac{\left( {I_{E} - x_{2}} \right)}{x_{1}}} & {{Eq}.\mspace{14mu} 10}\end{matrix}$where

Gest represents the estimated glucose concentration;

I_(E) is the signal measured at about 2.5 seconds;

x₁ is the slope (e.g., x₁=1.3e01);

x₂ is the intercept (e.g., x₂=6.9e02). As previously noted, the term “e”as used throughout this disclosure refers to scientific notation; e.g.,6.9e02=6.9×10²=690.

From the estimated glucose, the glucose concentration can be determinedfrom:

$\begin{matrix}{G_{o} = \frac{\left( {I_{s} - x_{4}} \right)}{x_{3}}} & {{Eq}.\mspace{14mu} 11}\end{matrix}$where

G_(O) represents the glucose concentration;

I_(S) is the signal measured at a specified sampling time S from Table3;

x₃ is the slope (e.g., x₃=9.6); and

x₄ is the intercept (e.g., x₄=4.8e02). As previously noted, the term “e”as used throughout this disclosure represents scientific notation; e.g.,4.8e02=4.8×10²=480.

Although the method may specify only one sampling time point, the methodmay include sampling as many time points as required, such as, forexample, sampling the signal output continuously (e.g., at specifiedsampling time such as, every 1 milliseconds to 100 milliseconds) fromthe start of the test sequence until at least about 10 seconds after thestart and the results stored for processing near the end of the testsequence. In this variation, the sampled signal output at the specifiedsampling time (which may be different from the predetermined samplingtime point) is the value used to calculate the analyte concentration.

It is noted that in the preferred embodiments, the measurement of asignal output for the value that is somewhat proportional to analyte(e.g., glucose) concentration is performed prior to the estimation ofthe hematocrit. Alternatively, the hematocrit level can be estimatedprior to the measurement of the preliminary glucose concentration. Ineither case, the estimated glucose measurement GE is obtained byEquation 3.3 with I_(E) sampled at about one of 2.5 seconds or 5seconds, as in FIG. 6D(2), the physical characteristic (e.g., Hct) isobtained by Equation 4 and the glucose measurement G is obtained byusing the measured signal output ID at the designated sampling timepoint(s) (e.g., the measured signal output ID being sampled at 3.5seconds or 6.5 seconds) for the signal transient 1000.

Thus, as another benefit of the teaching provided herein, a method ofdemonstrating increased accuracy of a test strip is heretofore isachieved. This method involves providing for a batch of test strips,typically in a batch of at least about 1500 test strips (and up to amillion or more test strips in certain cases), introducing a referentialsample containing a referential concentration of an analyte to each ofthe batch of test strips to initiate a test sequence. The methodinvolves, reacting the analyte to cause a physical transformation of theanalyte with the reagent between the two electrodes, determining aphysical characteristic of the referential sample, estimating theanalyte concentration, sampling an electrical output of the referentialsample at a dictated time point during the test sequence defined by themeasured or estimated physical characteristic of the sample and theestimated analyte concentration, and calculating an analyteconcentration based on the dictated time such that at least 95% of theanalyte concentration values of the batch of test strips are within ±10%of the referential analyte concentration for the range of hematocritfrom about 30% to about 55%, shown here in FIG. 6D(3).

In FIG. 6D(3), experiments were performed to quantify the improvement inthe glucose measurements from the method noted above. The quantificationof the improvement can be shown by the “bias” at different levels ofhematocrit. The bias, which is an estimate of the relative error in theglucose measurement, was calculated for each glucose concentrationdetermined with the method described in this example.

As can be seen in FIG. 6D(3), most or virtually all of the glucoseconcentration using this technique are within about ±10% bias forhematocrit from about 30% hematocrit to about 55% hematocrit. This isbelieved to be a breakthrough as the prior technique of sampling theoutput signal transient at a fixed time point (typically at about one of2.5 seconds or 5 seconds from the start of test sequence) for thisspecific type of test strip generally fails to show any bias of lessthan ±20%.

By virtue of the descriptions and teachings provided herein, we wereable to devise a glucose test strip that has a substrate, a plurality ofelectrodes disposed on the substrate and connected to respectiveelectrode connectors. The test strip 100 includes at least a reagentdisposed on at least one of the plurality of electrodes, in which atleast one of the electrodes is configured to sense a physicalcharacteristic of fluid sample deposited on the at least one electrodeand at least another of the electrodes is configured to measure outputsignal from the sample upon application of input signal to the sample.Included with the test strip are instructions for use with a glucosemeter. The instructions include indicia embedded in an appropriatecommunication medium (e.g., paper, computer, internet, audio or visualmedium or the like) to a user to insert the electrode connectors of thetest strip to a test strip port of the glucose meter. The meterindicated for use with the glucose test strip includes a test strip portconnector configured to connect to respective electrode connectors of atest strip, and a microprocessor in electrical communication with thetest strip port connector to apply electrical signals or senseelectrical signals from a plurality of electrodes of the test stripconnected to the respective electrode connectors of the test stripduring a test sequence. The instructions further include indiciaembedded in an appropriate communication medium (e.g., paper, computer,internet, audio or visual medium or the like) to the user to deposit afluid sample proximate at least one of the plurality of electrodes sothat the microprocessor 300 is operable to: (a) apply a first signal tothe plurality of electrodes so that a physical characteristic of a fluidsample deposited on the plurality of electrodes is determined; (b)estimate an analyte concentration based on a predetermined sampling timepoint during a test sequence; and (c) apply a second signal to theplurality of electrodes at a sampling time point during the testsequence dictated by the determined physical characteristic so that ananalyte concentration is calculated from the second signal.

Similarly, we were able to devise a glucose test strip 100 that has asubstrate, a plurality of electrodes disposed on the substrate andconnected to respective electrode connectors. The test strip includes atleast a reagent disposed on at least one of the plurality of electrodes,in which at least one of the electrodes is configured to sense aphysical characteristic of fluid sample deposited on the at least oneelectrode and at least another of the electrodes is configured tomeasure output signal from the sample upon application of input signalto the sample. Included with the test strip are instructions for usewith a glucose meter. The instructions include indicia embedded in anappropriate communication medium (e.g., paper, computer, internet, audioor visual means and the like) to a user to insert the electrodeconnectors of the test strip to a test strip port of the glucose meter.The meter indicated for use with the glucose test strip includes a teststrip port connector configured to connect to respective electrodeconnectors of a test strip, and a microprocessor in electricalcommunication with the test strip port connector to apply electricalsignals or sense electrical signals from a plurality of electrodes ofthe test strip connected to the respective electrode connectors of thetest strip during a test sequence. The instructions further includeindicia to the user to deposit a fluid sample proximate at least one ofthe plurality of electrodes so that the microprocessor is operable to:(a) apply a first signal to the plurality of electrodes so that aspecific sampling time determined from a physical characteristic of afluid sample is derived, (b) apply a second signal to the plurality ofelectrodes, and (c) measure a signal output from one of the plurality ofelectrodes at the specified sampling time so that an analyteconcentration is determined.

Likewise, we were able to devise a glucose test strip that has asubstrate, a plurality of electrodes disposed on the substrate andconnected to respective electrode connectors. The test strip includes atleast a reagent disposed on at least one of the plurality of electrodes,in which at least one of the electrodes is configured to sense aphysical characteristic of fluid sample deposited on the at least oneelectrode and at least another of the electrodes is configured tomeasure output signal from the sample upon application of input signalto the sample. Included with the test strip are instructions for usewith a glucose meter. The instructions include indicia embedded in anappropriate communication medium (e.g., paper, computer, internet, audioor visual means and the like) to a user to insert the electrodeconnectors of the test strip to a test strip port of the glucose meter.The meter indicated for use with the glucose test strip includes a teststrip port connector configured to connect to respective electrodeconnectors of a test strip, and a microprocessor in electricalcommunication with the test strip port connector to apply electricalsignals or sense electrical signals from a plurality of electrodes ofthe test strip connected to the respective electrode connectors of thetest strip during a test sequence. The instructions further includeindicia to the user to deposit a fluid sample proximate at least one ofthe plurality of electrodes so that the microprocessor is operable to:(a) apply a first signal to the plurality of electrodes so that aspecified sampling time and a batch slope determined from a physicalcharacteristic of a fluid sample are derived, (b) apply a second signalto the plurality of electrodes, and (c) measure a signal output from oneof the plurality of electrodes at the specified sampling time so that ananalyte concentration is determined based on the measured signal at thespecified sampling time and the batch slope.

Similarly, applicant was able to devise a glucose test strip that has asubstrate, a plurality of electrodes disposed on the substrate andconnected to respective electrode connectors. The test strip includes atleast a reagent disposed on at least one of the plurality of electrodes,in which at least one of the electrodes is configured to sense aphysical characteristic of fluid sample deposited on the at least oneelectrode and at least another of the electrodes is configured tomeasure output signal from the sample upon application of input signalto the sample. Included with the test strip are instructions for usewith a glucose meter. The instructions include indicia embedded in anappropriate communication medium (e.g., paper, computer, internet, audioor visual means and the like) to a user to insert the electrodeconnectors of the test strip to a test strip port of the glucose meter.The meter indicated for use with the glucose test strip includes a teststrip port connector configured to connect to respective electrodeconnectors of a test strip, and a microprocessor in electricalcommunication with the test strip port connector to apply electricalsignals or sense electrical signals from a plurality of electrodes ofthe test strip connected to the respective electrode connectors of thetest strip during a test sequence. The instructions further includeindicia to the user to deposit a fluid sample proximate at least one ofthe plurality of electrodes so that the microprocessor is operable to:(a) apply a first signal to the plurality of electrodes so that a batchslope defined by a physical characteristic of a fluid sample is derived,and (b) apply a second signal to the plurality of electrodes so that ananalyte concentration is determined based on the derived batch slope.

Although the techniques described herein have been directed todetermination of glucose, the techniques can also apply to otheranalytes (with appropriate modifications by those skilled in the art)that are affected by physical characteristic(s) of the fluid sample inwhich the analyte(s) is disposed in the fluid sample. For example, thephysical characteristic (e.g., hematocrit, viscosity or density and thelike) of a physiological fluid sample could be accounted for indetermination of ketone or cholesterol in the fluid sample, which may bephysiological fluid, calibration, or control fluid. Other biosensorconfigurations can also be utilized. For example, the biosensors shownand described in the following US Patents can be utilized with thevarious embodiments described herein: U.S. Pat. Nos. 6,179,979;6,193,873; 6,284,125; 6,413,410; 6,475,372; 6,716,577; 6,749,887;6,863,801; 6,890,421; 7,045,046; 7,291,256; 7,498,132, all of which areincorporated by reference in their entireties herein.

As is known, the detection of the physical characteristic does not haveto be done by alternating signals but can be done with other techniques.For example, a suitable sensor can be utilized (e.g., US PatentApplication Publication No. 20100005865 or EP1804048 B1) to determinethe viscosity or other physical characteristics. Alternatively, theviscosity can be determined and used to derive for hematocrits based onthe known relationship between hematocrits and viscosity as described in“Blood Rheology and Hemodynamics” by Oguz K. Baskurt, M.D., Ph.D., l andHerbert J. Meiselman, Sc.D., Seminars in Thrombosis and Hemostasis,volume 29, number 5, 2003.

As described earlier, the microcontroller or an equivalentmicroprocessor (and associated components that allow the microcontrollerto function for its intended purpose in the intended environment suchas, for example, the processor 300 in FIG. 2B) can be utilized withcomputer codes or software instructions to carry out the methods andtechniques described herein. Applicants note that the exemplarymicrocontroller 300 (along with suitable components for functionaloperation of the processor 300) in FIG. 2B is embedded with firmware orloaded with computer software representative of the logic diagrams inFIG. 6A(1), 6B(1); 6C(1); or 6D(1) and the microcontroller 300, alongwith associated connector 220 and interface 306 and equivalents thereof,are the means for: (a) determining a specified sampling time based on asensed or estimated physical characteristic, the specified sampling timebeing at least one time point or interval referenced from a start of atest sequence upon deposition of a sample on the test strip and (b)determining an analyte concentration based on the specified samplingtime. Alternatively, the means for determining may include means forapplying a first signal to the plurality of electrodes so that a batchslope defined by a physical characteristic of a fluid sample is derivedand for applying a second signal to the plurality of electrodes so thatan analyte concentration is determined based on the derived batch slopeand the specified sampling time. Furthermore, the means for determiningmay include means for estimating an analyte concentration based on apredetermined sampling time point from the start of the test sequenceand for selecting a specified sampling time from a matrix of estimatedanalyte concentration and sensed or estimated physical characteristic.Yet further, the means for determining may include means for selecting abatch slope based on the sensed or estimated physical characteristic andfor ascertaining the specified sampling time point from the batch slope.

A short discussion of the embodiments of the meter for the presentdisclosure is worthwhile at this point. In particular, hand-held testmeters for use with an analytical test strip in the determination of ananalyte (such as glucose) in a bodily fluid sample (i.e., a whole bloodsample) according to embodiments of the present disclosure include ahousing, a microcontroller block disposed in the housing, and aphase-shift-based hematocrit measurement block (also referred to as aphase-shift-based hematocrit circuit). In such hand-held test meters,the phase-shift-based hematocrit measurement block includes a signalgeneration sub-block, a low pass filter sub-block, an analytical teststrip sample cell interface sub-block, a transimpedance amplifiersub-block, and a phase detector sub-block. In addition, thephase-shift-based hematocrit measurement block and microcontroller blockare configured to measure the phase shift of a bodily fluid sample in asample cell of an analytical test strip inserted in the hand-held testmeter and the microcontroller block is also configured to compute thehematocrit of the bodily fluid sample based on the measured phase shift.

Hand-held test meters according to embodiments of the present disclosureare beneficial in that they provide improved accuracy of analytedetermination (such as glucose determination) in whole blood samples bymeasuring the hematocrit of the whole blood sample and then employingthe measured hematocrit during analyte determination.

Once one skilled in the art is apprised of the present disclosure, he orshe will recognize that an example of a hand-held test meter that can bereadily modified as a hand-held test meter according to the presentdisclosure is the commercially available OneTouch® Ultra® 2 glucosemeter from LifeScan Inc. (Milpitas, Calif.). Additional examples ofhand-held test meters that can also be modified are found in U.S. PatentApplication Publications No's. 2007/0084734 (published on Apr. 19, 2007)and 2007/0087397 (published on Apr. 19, 2007) and in InternationalPublication Number WO2010/049669 (published on May 6, 2010), each ofwhich is hereby incorporated herein in full by reference.

FIG. 8 is a simplified depiction of a hand-held test meter 100 accordingto an embodiment of the present disclosure. FIG. 9 is a simplified blockdiagram of various blocks of hand-held test meter 100. FIG. 10 is asimplified combined block diagram of a phase-shift-based hematocritmeasurement block of hand-held test meter 100. FIG. 11 is a simplifiedannotated schematic diagram of a dual low pass filter sub-block ofhand-held test meter 100. FIG. 12 is a simplified annotated schematicdiagram of a transimpedance amplifier sub-block of hand-held test meter100. FIG. 13 is a simplified annotated schematic block diagram ofportions of a phase-shift-based hematocrit measurement block ofhand-held test meter 100.

Referring to FIGS. 8 through 13, hand-held test meter 100 includes adisplay 102, a plurality of user interface buttons 104, a strip portconnector 106, a USB interface 108, and a housing 110 (see FIG. 8).Referring to FIG. 9 in particular, hand-held test meter 100 alsoincludes a microcontroller block 112, a phase-shift-based hematocritmeasurement block 114, a display control block 116, a memory block 118and other electronic components (not shown) for applying a test voltageto analytical test strip (labeled T_(S) in FIG. 8), and also formeasuring an electrochemical response (e.g., plurality of test currentvalues) and determining an analyte based on the electrochemicalresponse. To simplify the current descriptions, the figures do notdepict all such electronic circuitry.

Display 102 can be, for example, a liquid crystal display or a bi-stabledisplay configured to show a screen image. An example of a screen imagemay include a glucose concentration, a date and time, an error message,and a user interface for instructing an end user how to perform a test.

Strip port connector 106 is configured to operatively interface with ananalytical test strip TS, such as an electrochemical-based analyticaltest strip configured for the determination of glucose in a whole bloodsample. Therefore, the analytical test strip is configured for operativeinsertion into strip port connector 106 and to operatively interfacewith phase-shift-based hematocrit measurement block 114 via, forexample, suitable electrical contacts.

USB Interface 108 can be any suitable interface known to one skilled inthe art. USB Interface 108 is essentially a passive component that isconfigured to power and provide a data line to hand-held test meter 100.

Once an analytical test strip is interfaced with hand-held test meter100, or prior thereto, a bodily fluid sample (e.g., a whole bloodsample) is introduced into a sample chamber of the analytical teststrip. The analytical test strip can include enzymatic reagents thatselectively and quantitatively transform an analyte into anotherpredetermined chemical form. For example, the analytical test strip caninclude an enzymatic reagent with ferricyanide and glucose oxidase sothat glucose can be physically transformed into an oxidized form.

Memory block 118 of hand-held test meter 100 includes a suitablealgorithm and can be configured, along with microcontroller block 112 todetermine an analyte based on the electrochemical response of analyticaltest strip and the hematocrit of the introduced sample. For example, inthe determination of the analyte blood glucose, the hematocrit can beused to compensate for the effect of hematocrit on electrochemicallydetermined blood glucose concentrations.

Microcontroller block 112 is disposed within housing 110 and can includeany suitable microcontroller and/or micro-processer known to those ofskill in the art. One such suitable microcontroller is a microcontrollercommercially available from Texas Instruments, Dallas, Tex. USA and partnumber MSP430F5138. This microcontroller can generate a square wave of25 to 250 kHz and a 90-degree phase-shifted wave of the same frequencyand, thereby, function as a signal generation s-block described furtherbelow. MSP430F5138 also has Analog-to-Digital (A/D) processingcapabilities suitable for measuring voltages generated by phase shiftbased hematocrit measurement blocks employed in embodiments of thepresent disclosure.

Referring in particular to FIG. 10, phase-shift-based hematocritmeasurement block 114 includes a signal generation sub-block 120, a lowpass filter sub-block 122, an analytical test strip sample cellinterface sub-block 124, an optional calibration load block 126 (withinthe dashed lines of FIG. 10), a transimpedance amplifier sub-block 128,and a phase detector sub-block 130.

As described further below, phase-shift-based hematocrit measurementblock 114 and microcontroller block 112 are configured to measure thephase shift of a bodily fluid sample in a sample cell of an analyticaltest strip inserted in the hand-held test meter by, for example,measuring the phase shift of one or more high frequency electricalsignals driven through the bodily fluid sample. In addition,microcontroller block 112 is configured to compute the hematocrit of thebodily fluid based on the measured phase shift. Microcontroller 112 cancompute the hematocrit by, for example, employing an A/D converter tomeasure voltages received from a phase-detector sub-block, convert thevoltages into a phase-shift and then employing a suitable algorithm orlook-up table to convert the phase-shift into a hematocrit value. Onceapprised of the present disclosure, one skilled in the art willrecognize that such an algorithm and/or look-up table will be configuredto take into account various factors such as strip geometry (includingelectrode area and sample chamber volume) and signal frequency.

It has been determined that a relationship exists between the reactanceof a whole blood sample and the hematocrit of that sample. Electricalmodeling of a bodily fluid sample (i.e., a whole blood sample) asparallel capacitive and resistive components indicates that when analternating current (AC) signal is forced through the bodily fluidsample, the phase shift of the AC signal will be dependent on both thefrequency of the AC voltage and the hematocrit of the sample. Moreover,modeling indicates that hematocrit has a relatively minor effect on thephase shift when the frequency of the signal is in the range ofapproximately 10 kHz to 25 kHz and a maximum effect on the phase shiftwhen the frequency of the signal is in the range of approximately 250kHz to 500 KHz. Therefore, the hematocrit of a bodily fluid sample canbe measured by, for example, driving AC signals of known frequencythrough the bodily fluid sample and detecting their phase shift. Forexample, the phase-shift of a signal with a frequency in the range of 10kHz to 25 kHz can be used as a reference reading in such a hematocritmeasurement while the phase shift of a signal with a frequency in therange of 250 kHz to 500 kHz can be used as the primary measurement.

Referring to FIGS. 10 through 13 in particular, signal generationsub-block 120 can be any suitable signal generation block and isconfigured to generate a square wave (OV to Vref) of a desiredfrequency. Such a signal generation sub-block can, if desired, beintegrated into microcontroller block 112.

The signal generated by signal generation sub-block 120 is communicatedto dual low pass filter sub-block 122, which is configured to convertthe square wave signal to a sine wave signal of a predeterminedfrequency. The dual LPF of FIG. 11 is configured to provide both asignal of a first frequency (such as a frequency in the range of 10 kHzto 25 kHz) and a signal of a second frequency (such as a frequency inthe range of 250 kHz to 500 kHz) to the analytical test strip samplecell interface sub-block and an analytical test strips' sample chamber(also referred to as the HCT measurement cell). Selection of the firstand second frequency is accomplished using switch IC7 of FIG. 11. Thedual LPF of FIG. 11 includes employs two suitable operational amplifiers(IC4 and IC5) such as the operational amplifier available from TexasInstruments, Dallas, Tex., USA as high-speed, voltage feedback, CMOSoperational amplifier part number OPA354.

Referring to FIG. 11, F-DRV represents a square wave input of either alow or high frequency (e.g., 25 kHz or 250 kHz) and is connected to bothIC4 and IC5. Signal Fi-HIGH/LOW (from the microcontroller) selects theoutput of dual low pass filter sub-block 122 via switch IC7. C5 in FIG.11 is configured to block the operating voltage of dual low pass filtersub-block 122 from the HCT measurement cell.

Although a specific dual LPF is depicted in FIG. 11, dual low passfilter sub-block 122 can be any suitable low pass filter sub-block knownto one skilled in the art including, for example, any suitable multiplefeedback low pass filter, or a Sallen and Key low pass filter.

The sine wave produced by low pass filter sub-block 122 is communicatedto analytical test strip sample cell interface sub-block 124 where it isdriven across the sample cell of the analytical test strip (alsoreferred to as an HCT measurement cell). Analytical test strip samplecell interface block 124 can be any suitable sample cell interface blockincluding, for example, an interface block configured to operativelyinterface with the sample cell of the analytical test strip via firstelectrode and second electrodes of the analytical test strip disposed inthe sample cell. In such a configuration, the signal can be driven intothe sample cell (from the low pass filter sub-block) via the firstelectrode and picked-up from the sample cell (by the transimpedanceamplifier sub-block) via the second electrode as depicted in FIG. 13.

The current produced by driving the signal across the sample cell ispicked-up by transimpedance amplifier sub-block 128 and converted into avoltage signal for communication to phase detector sub-block 130.

Transimpedance sub-block 128 can be any suitable transimpedancesub-block known to one skilled in the art. FIG. 12 is a simplifiedannotated schematic block diagram of one such transimpedance amplifiersub-block (based on two OPA354 operational amplifiers, IC3 and IC9). Thefirst stage of TIA sub-block 128 operates at, for example, 400 mV, whichlimits the AC amplitude to +/−400 mV. The second stage of TIA sub-block128 operates at Vref/2, a configuration which enables the generation ofan output of the full span of the microcontroller A/D inputs. C9 of TIAsub-block 128 serves as a blocking component that only allows an AC sinewave signal to pass.

Phase detector sub-block 130 can be any suitable phase detectorsub-block that produces either a digital frequency that can be read backby microcontroller block 112 using a capture function, or an analogvoltage that can be read back by microcontroller block 112 using ananalog to digital converter. FIG. 13 depicts a schematic that includestwo such phase detector sub-blocks, namely an XOR phase detector (in theupper half of FIG. 13 and including IC22 and IC23) and a QuadratureDEMUX phase detector (in the lower half of FIG. 13 and including IC12and IC13).

FIG. 13 also depicts a calibration load sub-block 126 that includes aswitch (IC16) and a dummy load R7 and C6. Calibration load sub-block 126is configured for the dynamic measurement of a phase offset for theknown phase shift of zero degrees produced by resistor R7, thusproviding a phase offset for use in calibration. C6 is configured toforce a predetermined slight phase shift, e.g. to compensate for phasedelays caused by parasitic capacities in the signal traces to the samplecell, or for phase delays in the electrical circuits (LPF and TIA).

The Quadrature DEMUX phase detector circuit of FIG. 13 includes twoportions, one portion for a resistive part of the incoming AC signal andone portion for the reactive portion of the incoming AC signal. Use ofsuch two portions enables the simultaneous measurement of both theresistive and reactive portion of the AC signal and a measurement rangethat covers 0 degrees to 360 degrees. The Quadrature DEMUX circuit ofFIG. 13 generates two separate output voltages. One of these outputvoltages represents the “in phase measurement” and is proportional tothe “resistive” part of the AC signal, the other output voltagerepresents the “Quadrature Measurement” and is proportional to the“reactive part of the signal. The phase shift is calculated as:ϕ=tan⁻¹(V _(QUAD-PHASE) /V _(IN-PHASE))

Such a Quadrature DEMUX phase detector circuit can also be employed tomeasure the impedance of a bodily fluid sample in the sample cell. It ishypothesized, without being bound, that the impedance could be employedalong with the phase-shift, or independently thereof, to determine thehematocrit of the bodily sample. The amplitude of a signal forcedthrough the sample cell can be calculated using the two voltage outputsof the Quadrature DEMUX circuit as follows:Amplitude=SQR((V _(QUAD-PHASE))²+(V _(IN-PHASE))²)

This amplitude can then be compared to an amplitude measured for theknown resistor of calibration load block 126 to determine the impedance.

The XOR phase detector portion has a measurement range of 0° to 180°, oralternatively a measurement range of −90° to +90°, depending whether the“Square wave input from μC” is in phase to the sine wave or is set to a90° phase shift. The XOR phase detector produces an output frequencythat is always double the input frequency, however the duty cyclevaries. If both inputs are perfectly in phase, the output is LOW, ifboth inputs are 180° shifted the output is always HIGH. By integratingthe output signal (e.g. via a simple RC element) a voltage can begenerated that is directly proportional to the phase shift between bothinputs.

Once apprised of the present disclosure, one skilled in the art willrecognize that phase detector sub-blocks employed in embodiments of thepresent disclosure can take any suitable form and include, for example,forms that employ rising edge capture techniques, dual edge capturetechniques, XOR techniques and synchronous demodulation techniques.

Since low pass filter sub-block 122, transimpedance amplifier sub-block128 and phase detector sub-block 130 can introduce a residual phaseshift into phase-shift-based hematocrit measurement block 114,calibration load block 126 can be optionally included in thephase-shift-based hematocrit measurement block. Calibration load block126 is configured to be essentially resistive in nature (for example a33 k-ohm load) and, therefore, induces no phase shift between excitationvoltage and generated current. Calibration load block 126 is configuredto be switched in across the circuit to give a “zero” calibrationreading. Once calibrated, the hand-held test meter can measure the phaseshift of a bodily fluid sample, subtract the “zero” reading to compute acorrected phase shift and subsequently compute the bodily samplehematocrit based on the corrected phase shift.

FIG. 14 is a flow diagram depicting stages in a method 200 for employinga hand-held test meter and analytical test strip (e.g., anelectrochemical-based analytical test strip). Method 200, at step 210,includes introducing a whole blood sample into a sample cell of theanalytical test strip.

At step 220, a phase shift of the whole blood sample in the sample cellis measured using a phase-shift-based measurement block and amicrocontroller block of a hand-held test meter. Method 200 furtherincludes computing the hematocrit of whole blood sample based on themeasured phase shift using the microcontroller block (see step 230 ofFIG. 14).

Moreover, while the invention has been described in terms of particularvariations and illustrative figures, those of ordinary skill in the artwill recognize that the invention is not limited to the variations orfigures described. In addition, where methods and steps described aboveindicate certain events occurring in certain order, it is intended thatcertain steps do not have to be performed in the order described but inany order as long as the steps allow the embodiments to function fortheir intended purposes. Therefore, to the extent there are variationsof the invention, which are within the spirit of the disclosure orequivalent to the inventions found in the claims, it is the intent thatthis patent will cover those variations as well.

The invention claimed is:
 1. A method of determining an analyteconcentration from a fluid sample having an analyte of interest with abiosensor having at least two electrodes and a reagent disposed on atleast one electrode of the at least two electrodes, the methodcomprising the steps of: a) depositing the fluid sample on any one ofthe at least two electrodes to start an analyte test sequence; b)applying a first signal to the fluid sample in order to derive ahematocrit level of the fluid sample; c) obtaining the hematocrit levelof the fluid sample; d) specifying a sampling time based on the obtainedhematocrit level; e) driving a second signal to the fluid sample,wherein the second signal is applied from the start of the analyte testsequence; f) measuring an output signal at the specified sampling timefrom at least one electrode of the at least two electrodes; and g) usinga processor, calculating the analyte concentration of the fluid samplebased on the measured output signal, wherein the biosensor is a teststrip and the specified sampling time is either: i) calculated using anempirically derived equation of the form:SpecifiedSamplingTime=x ₁ H ^(x) ² +x ₃ where “SpecifiedSamplingTime” isdesignated as a time point in seconds from the start of the testsequence at which to sample the output signal of the test strip, Hrepresents the hematocrit level of the sample expressed as wholepercentages; x₁ is 4.3×10⁵ (s)+/−10%; x₂ is −3.9+/−10%; and x₃ is 4.8(s)+/−10%, or ii) the specified sampling time is estimated using astored look-up Table 1 based on the hematocrit level, expressed as wholepercentages as follows: TABLE 1 Hematocrit Level Sampling Time (T) (%)(seconds) 30 5.56 31 5.46 32 5.38 33 5.32 34 5.26 35 5.2 36 5.16 37 5.1238 5.08 39 5.06 40 5.02 41 5 42 5 43 4.98 44 4.96 45 4.96 46 4.94 474.92 48 4.92 49 4.9 50 4.9 51 4.9 52 4.88 53 4.88 54 4.88 55 4.86

and wherein the analyte is glucose and in which the method furthercomprises the step of annunciating the analyte concentration of thefluid sample.
 2. The method of claim 1, in which the steps of applyingthe first signal and the driving of the second signal is in sequentialorder.
 3. The method of claim 1, in which the step of applying of thefirst signal overlaps with the step of driving of the second signal. 4.The method of claim 1, in which the step of applying of the first signalcomprises the steps of directing an alternating signal to the sample sothat the hematocrit level of the fluid sample is determined from anoutput of the alternating signal.
 5. The method of claim 4, in which thestep of directing comprises the step of driving first and secondalternating signals at different respective frequencies in which a firstfrequency is lower than a second frequency.
 6. The method of claim 5, inwhich the first frequency is at least one order of magnitude lower thanthe second frequency.
 7. The method of claim 6, in which the firstfrequency comprises any frequency in the range of 10 kHz to 250 kHz. 8.The method of claim 1, in which the step of calculating of the analyteconcentration is computed using the processor with an equation of theform:$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{Slope} \right\rbrack$where G₀ represents the analyte concentration of the fluid sample; I_(E)represents, or is, a signal value or measurement proportional to analyteconcentration measured at the SpecifiedSamplingTime, quoted in nA; Sloperepresents the value obtained from calibration testing of a batch oftest strips of which this particular strip comes from, quoted innA(mg/dL); and Intercept represents the value obtained from calibrationtesting of a batch of test strips of which this particular strip comesfrom, quoted in nA.
 9. The method according to claim 1, the methodfurther comprising the steps of: using the processor, deriving a batchslope for the biosensor based on the hematocrit level from the obtainingstep, wherein the calculating of the analyte concentration is based onthe measured output signal at the specified sampling time and thederived batch slope, and wherein the biosensor is a test strip from abatch of test strips and the batch slope is defined as the measured orderived gradient of the line of best fit for a graph of measured glucoseconcentration plotted against actual glucose concentration for the batchof test strips.
 10. The method of claim 9, in which the derived batchslope is determined using the processor from an empirically derivedequation of the form:NewSlope=aH ² bH+c where NewSlope represents the derived batch slope(measured in arbitrary units); H is the measured or estimated hematocritlevel of the fluid sample (measured as percent); a is 1.4×10⁻⁶+/−10%, bis −3.8×10⁻⁴+/−10%, c is 3.6×10⁻²+/−10%.
 11. The method of claim 10, inwhich the step of calculating the analyte concentration is computed withan equation of the form:$G_{0} = \left\lbrack \frac{I_{E} - {Intercept}}{NewSlope} \right\rbrack$where G₀ represents the analyte concentration; I_(E) represents a signalproportional to analyte concentration measured at theSpecifiedSamplingTime, quoted in nA; NewSlope represents the valuederived from the measured or estimated physical characteristic; andIntercept represents the value obtained from calibration testing of thebatch of test strips of which this particular strip comes from, quotedin nA.
 12. A glucose meter comprising: a housing; a test strip portconnector configured to connect to the respective electrode connectorsof the test strip; and the processor configured to perform steps b) tog) of the method of claim
 1. 13. The glucose meter of claim 12, whereinthe processor is further programmed to determine the analyteconcentration of the sample based on the specific sampling time within10 seconds of a start of the analyte test sequence.
 14. A glucosemeasurement system comprising: a test strip including: a substrate; anda plurality of electrodes connected to respective electrode connectors;and the glucose meter according to claim 12.